WorldWideScience

Sample records for binary sequence generators

  1. Next-Generation Sequencing for Binary Protein-Protein Interactions

    Directory of Open Access Journals (Sweden)

    Bernhard eSuter

    2015-12-01

    Full Text Available The yeast two-hybrid (Y2H system exploits host cell genetics in order to display binary protein-protein interactions (PPIs via defined and selectable phenotypes. Numerous improvements have been made to this method, adapting the screening principle for diverse applications, including drug discovery and the scale-up for proteome wide interaction screens in human and other organisms. Here we discuss a systematic workflow and analysis scheme for screening data generated by Y2H and related assays that includes high-throughput selection procedures, readout of comprehensive results via next-generation sequencing (NGS, and the interpretation of interaction data via quantitative statistics. The novel assays and tools will serve the broader scientific community to harness the power of NGS technology to address PPI networks in health and disease. We discuss examples of how this next-generation platform can be applied to address specific questions in diverse fields of biology and medicine.

  2. METHOD OF GENERATING COMMON CRYPTOGRAPHIC KEYS FOR LOOSLY COINCIDENT BINARY SEQUENCES

    Directory of Open Access Journals (Sweden)

    V. L. Pivovarov

    2016-01-01

    Full Text Available The method of forming a common secret binary sequence between using an open communication channel is considered. The method is not based on common unidirectional functions and results in iterative elimination of distinct bits in the initial binary sequences with a certain percentage of mismatches, intentionally made by subscribers themselves. The cryptanalysis technique of this method based on the use of the deviation of aprior distribution of probabilities of inverting bits in the original binary sequences of subscribers from uniform distribution is proposed. Part of the bits in the final secret sequence can be identified accurately enough.

  3. Low autocorrelation binary sequences

    Science.gov (United States)

    Packebusch, Tom; Mertens, Stephan

    2016-04-01

    Binary sequences with minimal autocorrelations have applications in communication engineering, mathematics and computer science. In statistical physics they appear as groundstates of the Bernasconi model. Finding these sequences is a notoriously hard problem, that so far can be solved only by exhaustive search. We review recent algorithms and present a new algorithm that finds optimal sequences of length N in time O(N {1.73}N). We computed all optimal sequences for N≤slant 66 and all optimal skewsymmetric sequences for N≤slant 119.

  4. PERIODIC COMPLEMENTARY BINARY SEQUENCE PAIRS

    Institute of Scientific and Technical Information of China (English)

    XuChengqian; ZhaoXiaoqun

    2002-01-01

    A new set of binary sequences-Periodic Complementary Binary Sequence Pair (PCSP)is proposed .A new class of block design-Difference Family Pair (DFP)is also proposed .The relationship between PCSP and DFP,the properties and exising conditions of PCSP and the recursive constructions for PCSP are given.

  5. PERIODIC COMPLEMENTARY BINARY SEQUENCE PAIRS

    Institute of Scientific and Technical Information of China (English)

    Xu Chengqian; Zhao Xiaoqun

    2002-01-01

    A new set of binary sequences-Periodic Complementary Binary Sequence Pair (PCSP) is proposed. A new class of block design-Difference Family Pair (DFP) is also proposed.The relationship between PCSP and DFP, the properties and existing conditions of PCSP and the recursive constructions for PCSP are given.

  6. Bayesian analysis of binary sequences

    Science.gov (United States)

    Torney, David C.

    2005-03-01

    This manuscript details Bayesian methodology for "learning by example", with binary n-sequences encoding the objects under consideration. Priors prove influential; conformable priors are described. Laplace approximation of Bayes integrals yields posterior likelihoods for all n-sequences. This involves the optimization of a definite function over a convex domain--efficiently effectuated by the sequential application of the quadratic program.

  7. A measurement of disorder in binary sequences

    Science.gov (United States)

    Gong, Longyan; Wang, Haihong; Cheng, Weiwen; Zhao, Shengmei

    2015-03-01

    We propose a complex quantity, AL, to characterize the degree of disorder of L-length binary symbolic sequences. As examples, we respectively apply it to typical random and deterministic sequences. One kind of random sequences is generated from a periodic binary sequence and the other is generated from the logistic map. The deterministic sequences are the Fibonacci and Thue-Morse sequences. In these analyzed sequences, we find that the modulus of AL, denoted by |AL | , is a (statistically) equivalent quantity to the Boltzmann entropy, the metric entropy, the conditional block entropy and/or other quantities, so it is a useful quantitative measure of disorder. It can be as a fruitful index to discern which sequence is more disordered. Moreover, there is one and only one value of |AL | for the overall disorder characteristics. It needs extremely low computational costs. It can be easily experimentally realized. From all these mentioned, we believe that the proposed measure of disorder is a valuable complement to existing ones in symbolic sequences.

  8. Permutation Entropy for Random Binary Sequences

    Directory of Open Access Journals (Sweden)

    Lingfeng Liu

    2015-12-01

    Full Text Available In this paper, we generalize the permutation entropy (PE measure to binary sequences, which is based on Shannon’s entropy, and theoretically analyze this measure for random binary sequences. We deduce the theoretical value of PE for random binary sequences, which can be used to measure the randomness of binary sequences. We also reveal the relationship between this PE measure with other randomness measures, such as Shannon’s entropy and Lempel–Ziv complexity. The results show that PE is consistent with these two measures. Furthermore, we use PE as one of the randomness measures to evaluate the randomness of chaotic binary sequences.

  9. What's Next? Judging Sequences of Binary Events

    Science.gov (United States)

    Oskarsson, An T.; Van Boven, Leaf; McClelland, Gary H.; Hastie, Reid

    2009-01-01

    The authors review research on judgments of random and nonrandom sequences involving binary events with a focus on studies documenting gambler's fallacy and hot hand beliefs. The domains of judgment include random devices, births, lotteries, sports performances, stock prices, and others. After discussing existing theories of sequence judgments,…

  10. What's Next? Judging Sequences of Binary Events

    Science.gov (United States)

    Oskarsson, An T.; Van Boven, Leaf; McClelland, Gary H.; Hastie, Reid

    2009-01-01

    The authors review research on judgments of random and nonrandom sequences involving binary events with a focus on studies documenting gambler's fallacy and hot hand beliefs. The domains of judgment include random devices, births, lotteries, sports performances, stock prices, and others. After discussing existing theories of sequence judgments,…

  11. Debris disks in main sequence binary systems

    CERN Document Server

    Trilling, D E; Stapelfeldt, K R; Rieke, G H; Su, K Y L; Gray, R O; Corbally, C J; Bryden, G; Chen, C H; Boden, A; Beichman, C A

    2006-01-01

    We observed 69 A3-F8 main sequence binary star systems using the Multiband Imaging Photometer for Spitzer onboard the Spitzer Space Telescope. We find emission significantly in excess of predicted photospheric flux levels for 9(+4/-3)% and 40(+7/-6)% of these systems at 24 and 70 microns, respectively. Twenty two systems total have excess emission, including four systems that show excess emission at both wavelengths. A very large fraction (nearly 60%) of observed binary systems with small (<3 AU) separations have excess thermal mission. We interpret the observed infrared excesses as thermal emission from dust produced by collisions in planetesimal belts. The incidence of debris disks around main sequence A3-F8 binaries is marginally higher than that for single old AFGK stars. Whatever combination of nature (birth conditions of binary systems) and nurture (interactions between the two stars) drives the evolution of debris disks in binary systems, it is clear that planetesimal formation is not inhibited to a...

  12. Minimum degree and density of binary sequences

    DEFF Research Database (Denmark)

    Brandt, Stephan; Müttel, J.; Rautenbach, D.

    2010-01-01

    For d,k∈N with k ≤ 2d, let g(d,k) denote the infimum density of binary sequences (x)∈{0,1} which satisfy the minimum degree condition σ(x+) ≥ k for all i∈Z with xi=1. We reduce the problem of computing g(d,k) to a combinatorial problem related to the generalized k-girth of a graph G which...

  13. Next-generation sequencing

    DEFF Research Database (Denmark)

    Rieneck, Klaus; Bak, Mads; Jønson, Lars

    2013-01-01

    the feasibility of predicting the fetal KEL1 phenotype using next-generation sequencing (NGS) technology. STUDY DESIGN AND METHODS: The KEL1/2 single-nucleotide polymorphism was polymerase chain reaction (PCR) amplified with one adjoining base, and the PCR product was sequenced using a genome analyzer (GAIIx......, Illumina); several millions of PCR sequences were analyzed. RESULTS: The results demonstrated the feasibility of diagnosing the fetal KEL1 or KEL2 blood group from cell-free DNA purified from maternal plasma. CONCLUSION: This method requires only one primer pair, and the large amount of sequence...

  14. Fast algorithms for generating binary holograms

    CERN Document Server

    Stuart, Dustin; Kuhn, Axel

    2014-01-01

    We describe three algorithms for generating binary-valued holograms. Our methods are optimised for producing large arrays of tightly focussed optical tweezers for trapping particles. Binary-valued holograms allow us to use a digital mirror device (DMD) as the display element, which is much faster than other alternatives. We describe how our binary amplitude holograms can be used to correct for phase errors caused by optical aberrations. Furthermore, we compare the speed and accuracy of the algorithms for both periodic and arbitrary arrangements of traps, which allows one to choose the ideal scheme depending on the circumstances.

  15. Entropies of short binary sequences in heart period dynamics.

    Science.gov (United States)

    Cysarz, D; Bettermann, H; van Leeuwen, P

    2000-06-01

    Dynamic aspects of R-R intervals have often been analyzed by means of linear and nonlinear measures. The goal of this study was to analyze binary sequences, in which only the dynamic information is retained, by means of two different aspects of regularity. R-R interval sequences derived from 24-h electrocardiogram (ECG) recordings of 118 healthy subjects were converted to symbolic binary sequences that coded the beat-to-beat increase or decrease in the R-R interval. Shannon entropy was used to quantify the occurrence of short binary patterns (length N = 5) in binary sequences derived from 10-min intervals. The regularity of the short binary patterns was analyzed on the basis of approximate entropy (ApEn). ApEn had a linear dependence on mean R-R interval length, with increasing irregularity occurring at longer R-R interval length. Shannon entropy of the same sequences showed that the increase in irregularity is accompanied by a decrease in occurrence of some patterns. Taken together, these data indicate that irregular binary patterns are more probable when the mean R-R interval increases. The use of surrogate data confirmed a nonlinear component in the binary sequence. Analysis of two consecutive 24-h ECG recordings for each subject demonstrated good intraindividual reproducibility of the results. In conclusion, quantification of binary sequences derived from ECG recordings reveals properties that cannot be found using the full information of R-R interval sequences.

  16. Binary Sequences from a Pair of Elliptic Curves

    Institute of Scientific and Technical Information of China (English)

    CHEN Zhixiong; ZHANG Ning; XIAO Guozhen

    2006-01-01

    A family of binary sequences were constructed by using an elliptic curve and its twisted curves over finite fields. It was shown that these sequences possess "good" cryptographic properties of 0-1 distribution, long period and large linear complexity. The results indicate that such sequences provide strong potential applications in cryptography.

  17. Next-generation sequencing

    DEFF Research Database (Denmark)

    Rieneck, Klaus; Bak, Mads; Jønson, Lars

    2013-01-01

    information obtained allows well for statistical analysis of the data. This general approach can be integrated into current laboratory practice and has numerous applications. Besides DNA-based predictions of blood group phenotypes, platelet phenotypes, or sickle cell anemia, and the determination of zygosity......, Illumina); several millions of PCR sequences were analyzed. RESULTS: The results demonstrated the feasibility of diagnosing the fetal KEL1 or KEL2 blood group from cell-free DNA purified from maternal plasma. CONCLUSION: This method requires only one primer pair, and the large amount of sequence...

  18. The r-step Fibonacci-Hurwitz sequences in the binary polyhedral group

    Science.gov (United States)

    Deveci, Ömür; Ćiçekci, Deniz

    2016-04-01

    In [1], Deveci et al. defined the r-step Fibonacci-Hurwitz sequences from the Hurwitz matrices obtained from the characteristic polynomial of the k-step Fibonacci sequence. Also, they extended the r-step Fibonacci-Hurwitz sequences to groups. In this work, we obtain the periods of the r-step Fibonacci-Hurwitz sequences in the binary polyhedral group for generating triple {x, y, z} and generating pair {y, z} by the aid of the periods of the r-step Fibonacci-Hurwitz sequences according to modulo m.

  19. Next-Generation Sequencing Platforms

    Science.gov (United States)

    Mardis, Elaine R.

    2013-06-01

    Automated DNA sequencing instruments embody an elegant interplay among chemistry, engineering, software, and molecular biology and have built upon Sanger's founding discovery of dideoxynucleotide sequencing to perform once-unfathomable tasks. Combined with innovative physical mapping approaches that helped to establish long-range relationships between cloned stretches of genomic DNA, fluorescent DNA sequencers produced reference genome sequences for model organisms and for the reference human genome. New types of sequencing instruments that permit amazing acceleration of data-collection rates for DNA sequencing have been developed. The ability to generate genome-scale data sets is now transforming the nature of biological inquiry. Here, I provide an historical perspective of the field, focusing on the fundamental developments that predated the advent of next-generation sequencing instruments and providing information about how these instruments work, their application to biological research, and the newest types of sequencers that can extract data from single DNA molecules.

  20. Quasiequilibrium sequences of binary strange quark stars in general relativity

    CERN Document Server

    Limousin, F; Gourgoulhon, E; Limousin, Francois; Gondek-Rosinska, Dorota; Gourgoulhon, Eric

    2004-01-01

    Inspiraling compact binaries are expected to be the strongest sources of gravitational waves for VIRGO, LIGO and other laser interferometers. We present the first computations of quasi-equilibrium sequences of compact binaries containing two strange quark stars (which are currently considered as a possible alternative to neutron stars). We study a precoalescing stage in the conformal flatness approximation of general relativity using a multidomain spectral method. A hydrodynamical treatment is performed under the assumption that the flow is irrotational.

  1. Independence divergence-generated binary trees of amino acids.

    Science.gov (United States)

    Tusnády, G E; Tusnády, G; Simon, I

    1995-05-01

    The discovery of the relationship between amino acids is important in terms of the replacement ability, as used in protein engineering homology studies, and gaining a better understanding of the roles which various properties of the residues play in the creation of a unique, stable, 3-D protein structure. Amino acid sequences of proteins edited by evolution are anything but random. The measure of nonrandomness, i.e. the level of editing, can be characterized by an independence divergence value. This parameter is used to generate binary tree relationships between amino acids. The relationships of residues presented in this paper are based on protein building features and not on the physico-chemical characteristics of amino acids. This approach is not biased by the tautology present in all sequence similarity-based relationship studies. The roles which various physico-chemical characteristics play in the determination of the relationships between amino acids are also discussed.

  2. On the Eccentricity Excitation in Post-main-sequence Binaries

    Science.gov (United States)

    Rafikov, Roman R.

    2016-10-01

    Several classes of stellar binaries with post-main-sequence (post-MS) components—millisecond pulsars with the white dwarf companions (MSP+WD) and periods of {P}b∼ 30 days, binaries hosting post-asymptotic giant branch stars, or barium stars with {P}b ∼ several years—feature high eccentricities (up to 0.4) despite the expectation of their efficient tidal circularization during their post-MS evolution. It was suggested that the eccentricities of these binaries can be naturally excited by their tidal coupling to the circumbinary disk, formed by the material ejected from the binary. Here we critically reassess this idea using simple arguments rooted in the global angular momentum conservation of the disk+binary system. Compared to previous studies, we (1) fully account for the viscous spreading of the circumbinary disk, (2) consider the possibility of reaccretion from the disk onto the binary (in agreement with simulations and empirical evidence), and (3) allow for the reduced viscosity after the disk expands, cools, and forms dust. These ingredients conspire to significantly lower the efficiency of eccentricity excitation by the disk tides. We find that explaining eccentricities of the post-MS binaries is difficult and requires massive (≳ {10}-2 {M}ȯ ), long-lived (≳ {10}5 years) circumbinary disks that do not reaccrete. While disk tides may account for the eccentricities of the MSP+WD binaries, we show reaccretion to also be detrimental for these systems. Reduced efficiency of the disk-driven excitation motivates the study of alternative mechanisms for producing the peculiar eccentricities of the post-MS binaries.

  3. On the extendibility of partially and Markov exchangeable binary sequences

    CERN Document Server

    Di Cecco, Davide

    2009-01-01

    In [Fortini et al., Stoch. Proc. Appl. 100 (2002), 147--165] it is demonstrated that a recurrent Markov exchangeable process in the sense of Diaconis and Freedman is essentially a partially exchangeable process in the sense of de Finetti. In case of finite sequences there is not such an equivalence. We analyze both finite partially exchangeable and finite Markov exchangeable binary sequences and formulate necessary and sufficient conditions for extendibility in both cases.

  4. LSM is not generated by binary functions

    CERN Document Server

    McQuillan, Colin

    2011-01-01

    Bulatov et al. [1] defined the operation of (efficient) pps_\\omega-definability in order to study the computational complexity of certain approximate counting problems. They asked whether all log-supermodular functions can be defined by binary implication and unary functions in this sense. We give a negative answer to this question.

  5. Binary De Bruijn sequences for DS-CDMA systems: analysis and results

    Directory of Open Access Journals (Sweden)

    Spinsante Susanna

    2011-01-01

    Full Text Available Abstract Code division multiple access (CDMA using direct sequence (DS spread spectrum modulation provides multiple access capability essentially thanks to the adoption of proper sequences as spreading codes. The ability of a DS-CDMA receiver to detect the desired signal relies to a great extent on the auto-correlation properties of the spreading code associated to each user; on the other hand, multi-user interference rejection depends on the cross-correlation properties of all the spreading codes in the considered set. As a consequence, the analysis of new families of spreading codes to be adopted in DS-CDMA is of great interest. This article provides results about the evaluation of specific full-length binary sequences, the De Bruijn ones, when applied as spreading codes in DS-CDMA schemes, and compares their performance to other families of spreading codes commonly used, such as m-sequences, Gold, OVSF, and Kasami sequences. While the latter sets of sequences have been specifically designed for application in multi-user communication contexts, De Bruijn sequences come from combinatorial mathematics, and have been applied in completely different scenarios. Considering the similarity of De Bruijn sequences to random sequences, we investigate the performance resulting by applying them as spreading codes. The results herein presented suggest that binary De Bruijn sequences, when properly selected, may compete with more consolidated options, and encourage further investigation activities, specifically focused on the generation of longer sequences, and the definition of correlation-based selection criteria.

  6. OVERSAMPLED CHAOTIC BINARY SEQUENCES FOR MULTIPLE ACCESS COMMUNICATION

    Institute of Scientific and Technical Information of China (English)

    Zhang Hongtao; Wang Huiyun; Ding Runtao

    2002-01-01

    Noise interference and multiple access interference are the main impairment to the performance of DS/CDMA communication system. This letter presents that OverSampled Chaotic Map (OSCM) binary sequences are secure as spreading sequences, and based on the optimal quantizing method, the BER performance of the system has been derived in detail, the internal relationships among the number of users, the power of noise and the length of code chips are revealed in mathematical formulae. The performance of the system can be improved by employing these formulae. Numerical results conform the efficiency of discussion in this letter.

  7. Pseudo-random sequence generator based on the generalized Henon map

    Institute of Scientific and Technical Information of China (English)

    ZHENG Fan; TIAN Xiao-jian; SONG Jing-yi; LI Xue-yan

    2008-01-01

    By analysis and comparison of several chaotic systems that are applied to generate pseudo-random sequence, the generalized Henon map is proposed as a pseudo-random sequence generator. A new algorithm is created to solve the problem of non-uniform distribution of the sequence generated by the generalized Henon map. First, move the decimal point of elements in the sequence to the right; then, cut off the integer; and finally, quantify it into a binary sequence. Statistical test, security analysis, and the application of image encryption have strongly supported the good random statistical characteristics, high linear complexity, large key space, and great sensitivity of the binary sequence.

  8. Dynamic Test Generation for Large Binary Programs

    Science.gov (United States)

    2009-11-12

    Patrice Godefroid, Dennis Jeffries, and Adam Kiezun. The SAGE system builds on the iDNA/Time Travel Debugging and Nirvana infrastructure produced by... Nirvana [7] or Valgrind [74] (Catchconv is an example of the latter approach [70].) SAGE adopts offline trace-based constraint generation for two reasons

  9. A fast algorithm for determining the linear complexity of a binary sequence with period 2npm

    Institute of Scientific and Technical Information of China (English)

    魏仕民; 肖国镇; 陈钟

    2001-01-01

    An efficient algorithm for determining the linear complexity and the minimal polynomial of a binary sequence with period 2npm is proposed and proved, where 2 is a primitive root modulo p2. The new algorithm generalizes the algorithm for computing the linear complexity of a binary sequence with period 2 n and the algorithm for computing the linear complexity of a binary sequence with period pn,where 2 is a primitive root modulo p2.

  10. Binary interactions with high accretion rates onto main sequence stars

    Science.gov (United States)

    Shiber, Sagiv; Schreier, Ron; Soker, Noam

    2016-07-01

    Energetic outflows from main sequence stars accreting mass at very high rates might account for the powering of some eruptive objects, such as merging main sequence stars, major eruptions of luminous blue variables, e.g., the Great Eruption of Eta Carinae, and other intermediate luminosity optical transients (ILOTs; red novae; red transients). These powerful outflows could potentially also supply the extra energy required in the common envelope process and in the grazing envelope evolution of binary systems. We propose that a massive outflow/jets mediated by magnetic fields might remove energy and angular momentum from the accretion disk to allow such high accretion rate flows. By examining the possible activity of the magnetic fields of accretion disks, we conclude that indeed main sequence stars might accrete mass at very high rates, up to ≈ 10-2 M ⊙ yr-1 for solar type stars, and up to ≈ 1 M ⊙ yr-1 for very massive stars. We speculate that magnetic fields amplified in such extreme conditions might lead to the formation of massive bipolar outflows that can remove most of the disk's energy and angular momentum. It is this energy and angular momentum removal that allows the very high mass accretion rate onto main sequence stars.

  11. Composite Binary Sequences with a Large Ensemble and Zero Correlation Zone

    Directory of Open Access Journals (Sweden)

    S. S. Yudachev

    2015-01-01

    Full Text Available The article considers a proposed class of derived signals such as composite binary sequences for application in advanced spread spectrum radio systems of various purposes, using signals based on spectrum spreading by direct sequence method. Considered composite sequences, having a representative set of lengths and unique correlation properties, compares favorably with the widely used at present large ensembles formed on a single algorithmic basis. To evaluate the properties of the composite sequences generated on the basis of two components - the Barker code and Kerdock sequences, expressions of periodic and aperiodic correlation functions are given.An algorithm for generating practical ensembles of composite sequences is presented. On the basis of the algorithm and its software implementation in C #, the samples of the sequence ensembles of various lengths were obtained and their periodic and aperiodic correlation functions and statistical characteristics were studied in detail. As an illustration, some of the most typical correlation functions are presented. The most remarkable characteristics allowing a ssessing the feasibility of using this type of sequences in the design of specific types of radio systems are considered.On the basis of the proposed program and the performed calculations the conclusions can be drawn about the possibility of using the sequences of these classes, with the aim of reducing intra-system disturbance in the projected spread spectrum CDMA.

  12. THE CONSTRUCTIONS OF ALMOST BINARY SEQUENCE PAIRS WITH THREE-LEVEL CORRELATION BASED ON CYCLOTOMY

    Institute of Scientific and Technical Information of China (English)

    Peng Xiuping; Xu Chengqian

    2012-01-01

    In this paper,a new class of almost binary sequence pair with a single zero element is presented.The almost binary sequence pairs with three-level correlation are constructed based on cyclotomic numbers of order 2,4,and 6.Most of them have good correlation and balance property,whose maximum nontrivial correlation magnitudes are 2 and the difference between the numbers of occurrence of +1's and -1's are 0 or 1.In addition,the corresponding binary sequence pairs are investigated as well and we can also get some kinds of binary sequence pairs with optimum balance and good correlation.

  13. Discovering binary codes for documents by learning deep generative models.

    Science.gov (United States)

    Hinton, Geoffrey; Salakhutdinov, Ruslan

    2011-01-01

    We describe a deep generative model in which the lowest layer represents the word-count vector of a document and the top layer represents a learned binary code for that document. The top two layers of the generative model form an undirected associative memory and the remaining layers form a belief net with directed, top-down connections. We present efficient learning and inference procedures for this type of generative model and show that it allows more accurate and much faster retrieval than latent semantic analysis. By using our method as a filter for a much slower method called TF-IDF we achieve higher accuracy than TF-IDF alone and save several orders of magnitude in retrieval time. By using short binary codes as addresses, we can perform retrieval on very large document sets in a time that is independent of the size of the document set using only one word of memory to describe each document.

  14. Steady-state analysis of delay-locked loops tracking binary Markovian sequences

    Science.gov (United States)

    Nagata, Keisuke; Fujisaka, Hisato; Kamio, Takeshi; Ahn, Chang-Jun; Haeiwa, Kazuhisa

    We analyze stationary phase tracking error of delay-locked loops (DLL) in direct spread code division multiple access (DS-CDMA) using Markovian spreading sequences. The phase tracking error is caused by noise generated inside of DLLs by multiple access interferences. When binary Markovian sequences are used, the noise is not considered as white Gaussian noise. This makes analysis of the tracking error difficult. In this paper, we describe DLLs by stochastic difference equations and derive forward evolutional equations of the probability distribution of the states of DLLs. Applying path integral analysis to the evolutional equations, we obtained stationary distribution. We found from the distribution that Markovian spreading sequences with negative eigenvalue were effective in decreasing stationary phase tracking error of not only a type of DLL in asynchronous CDMA but also DLLs in chip-synchronous CDMA.

  15. Concept For Generation Of Long Pseudorandom Sequences

    Science.gov (United States)

    Wang, C. C.

    1990-01-01

    Conceptual very-large-scale integrated (VLSI) digital circuit performs exponentiation in finite field. Algorithm that generates unusually long sequences of pseudorandom numbers executed by digital processor that includes such circuits. Concepts particularly advantageous for such applications as spread-spectrum communications, cryptography, and generation of ranging codes, synthetic noise, and test data, where usually desirable to make pseudorandom sequences as long as possible.

  16. Design of Digital Hybrid Chaotic Sequence Generator

    Institute of Scientific and Technical Information of China (English)

    RAO Nini; ZENG Dong

    2004-01-01

    The feasibility of the hybrid chaotic sequences as the spreading codes in code divided multiple access(CDMA) system is analyzed.The design and realization of the digital hybrid chaotic sequence generator by very high speed integrated circuit hardware description language(VHDL) are described.A valid hazard canceledl method is presented.Computer simulations show that the stable digital sequence waveforms can be produced.The correlations of the digital hybrid chaotic sequences are compared with those of m-sequences.The results show that the correlations of the digital hybrid chaotic sequences are almost as good as those of m-sequences.The works in this paper explored a road for the practical applications of chaos.

  17. A new system identification approach to identify genetic variants in sequencing studies for a binary phenotype.

    Science.gov (United States)

    Kang, Guolian; Bi, Wenjian; Zhao, Yanlong; Zhang, Ji-Feng; Yang, Jun J; Xu, Heng; Loh, Mignon L; Hunger, Stephen P; Relling, Mary V; Pounds, Stanley; Cheng, Cheng

    2014-01-01

    We propose in this paper a set-valued (SV) system model, which is a generalized form of logistic (LG) and Probit (Probit) regression, to be considered as a method for discovering genetic variants, especially rare genetic variants in next-generation sequencing studies, for a binary phenotype. We propose a new SV system identification method to estimate all underlying key system parameters for the Probit model and compare it with the LG model in the setting of genetic association studies. Across an extensive series of simulation studies, the Probit method maintained type I error control and had similar or greater power than the LG method, which is robust to different distributions of noise: logistic, normal, or t distributions. Additionally, the Probit association parameter estimate was 2.7-46.8-fold less variable than the LG log-odds ratio association parameter estimate. Less variability in the association parameter estimate translates to greater power and robustness across the spectrum of minor allele frequencies (MAFs), and these advantages are the most pronounced for rare variants. For instance, in a simulation that generated data from an additive logistic model with an odds ratio of 7.4 for a rare single nucleotide polymorphism with a MAF of 0.005 and a sample size of 2,300, the Probit method had 60% power whereas the LG method had 25% power at the α = 10(-6) level. Consistent with these simulation results, the set of variants identified by the LG method was a subset of those identified by the Probit method in two example analyses. Thus, we suggest the Probit method may be a competitive alternative to the LG method in genetic association studies such as candidate gene, genome-wide, or next-generation sequencing studies for a binary phenotype.

  18. A New System Identification Approach to Identifying Genetic Variants in Sequencing Studies for A Binary Phenotype

    Science.gov (United States)

    Kang, Guolian; Bi, Wenjian; Zhao, Yanlong; Zhang, Ji-Feng; Yang, Jun J.; Xu, Heng; Loh, Mignon L.; Hunger, Stephen P.; Relling, Mary V.; Pounds, Stanley; Cheng, Cheng

    2014-01-01

    We propose in this paper a set-valued (SV) system model, which is a generalized form of Logistic (LG) and Probit (Probit) regression, to be considered as a method for discovering genetic variants, especially rare genetic variants in next generation sequencing studies, for a binary phenotype. We propose a new set-valued system identification method to estimate all the underlying key system parameters for the Probit model and compare it with the LG model in the setting of genetic association studies. Across an extensive series of simulation studies, the Probit method maintained Type I error control and had similar or greater power than the LG method which is robust to different distributions of noise: logistic, normal or t distributions. Additionally, the Probit association parameter estimate was 2.7–46.8 fold less variable than the LG log-odds ratio association parameter estimate. Less variability in the association parameter estimate translates to greater power and robustness across the spectrum of minor allele frequencies (MAFs), and these advantages are the most pronounced for rare variants. For instance, in a simulation that generated data from an additive logistic model with odds ratio of 7.4 for a rare single nucleotide polymorphism with a MAF of 0.005 and a sample size of 2300, the Probit method had 60% power whereas the LG method had 25% power at the α=10−6 level. Consistent with these simulation results, the set of variants identified by the LG method was a subset of those identified by the Probit method in two example analyses. Thus, we suggest the Probit method may be a competitive alternative to the LG method in genetic association studies such as candidate gene, genome-wide, or next generation sequencing studies for a binary phenotype. PMID:25096228

  19. Similarity Estimation Between DNA Sequences Based on Local Pattern Histograms of Binary Images

    Institute of Scientific and Technical Information of China (English)

    Yusei Kobori; Satoshi Mizuta

    2016-01-01

    Graphical representation of DNA sequences is one of the most popular techniques for alignment-free sequence comparison. Here, we propose a new method for the feature extraction of DNA sequences represented by binary images, by estimating the similarity between DNA sequences using the frequency histograms of local bitmap patterns of images. Our method shows linear time complexity for the length of DNA sequences, which is practical even when long sequences, such as whole genome sequences, are compared. We tested five distance measures for the estimation of sequence similarities, and found that the histogram intersection and Manhattan distance are the most appropriate ones for phylogenetic analyses.

  20. Rotational mixing in tidally locked massive main-sequence binaries

    CERN Document Server

    de Mink, S E; Langer, N; Pols, O R

    2008-01-01

    One of the main uncertainties in evolutionary calculations of massive stars is the efficiency of internal mixing. It changes the chemical profile inside the star and can therefore affect the structure and further evolution. We demonstrate that eclipsing binaries, in which the tides synchronize the rotation period of the stars and the orbital period, constitute a potentially strong test for the efficiency of rotational mixing. We present detailed stellar evolutionary models of massive binaries assuming the composition of the Small Magellanic Cloud. In these models we find enhancements in the surface nitrogen abundance of up to 0.6 dex.

  1. Generating large misalignments in gapped and binary discs

    Science.gov (United States)

    Owen, James E.; Lai, Dong

    2017-08-01

    Many protostellar gapped and binary discs show misalignments between their inner and outer discs; in some cases, ˜70° misalignments have been observed. Here, we show that these misalignments can be generated through a secular resonance between the nodal precession of the inner disc and the precession of the gap-opening (stellar or massive planetary) companion. An evolving protostellar system may naturally cross this resonance during its lifetime due to disc dissipation and/or companion migration. If resonance crossing occurs on the right time-scale, of the order of a few million years, characteristic for young protostellar systems, the inner and outer discs can become highly misaligned, with misalignments ≳ 60° typical. When the primary star has a mass of order a solar mass, generating a significant misalignment typically requires the companion to have a mass of ˜0.01-0.1 M⊙ and an orbital separation of tens of astronomical units. The recently observed companion in the cavity of the gapped, highly misaligned system HD 142527 satisfies these requirements, indicating that a previous resonance crossing event misaligned the inner and outer discs. Our scenario for HD 142527's misaligned discs predicts that the companion's orbital plane is aligned with the outer disc's; this prediction should be testable with future observations as the companion's orbit is mapped out. Misalignments observed in several other gapped disc systems could be generated by the same secular resonance mechanism.

  2. Pre-main sequence spectroscopic binaries suitable for VLTI observations

    CERN Document Server

    Guenther, E W; Mundt, R; Covino, E; Alcalá, J M; Cusano, F; Stecklum, B

    2007-01-01

    A severe problem of the research in star-formation is that the masses of young stars are almost always estimated only from evolutionary tracks. Since the tracks published by different groups differ, it is often only possible to give a rough estimate of the masses of young stars. It is thus crucial to test and calibrate the tracks. Up to now, only a few tests of the tracks could be carried out. However, with the VLTI it is now possible to set constrains on the tracks by determining the masses of many young binary stars precisely. In order to use the VLTI efficiently, a first step is to find suitable targets, which is the purpose of this work. Given the distance of nearby star-forming regions, suitable VLTI targets are binaries with orbital periods between at least 50 days, and few years. Although a number of surveys for detecting spectroscopic binaries have been carried out, most of the binaries found so far have periods which are too short. We thus surveyed the Chamaeleon, Corona Australis, Lupus, Sco-Cen, rh...

  3. Comparison of next-generation sequencing systems.

    Science.gov (United States)

    Liu, Lin; Li, Yinhu; Li, Siliang; Hu, Ni; He, Yimin; Pong, Ray; Lin, Danni; Lu, Lihua; Law, Maggie

    2012-01-01

    With fast development and wide applications of next-generation sequencing (NGS) technologies, genomic sequence information is within reach to aid the achievement of goals to decode life mysteries, make better crops, detect pathogens, and improve life qualities. NGS systems are typically represented by SOLiD/Ion Torrent PGM from Life Sciences, Genome Analyzer/HiSeq 2000/MiSeq from Illumina, and GS FLX Titanium/GS Junior from Roche. Beijing Genomics Institute (BGI), which possesses the world's biggest sequencing capacity, has multiple NGS systems including 137 HiSeq 2000, 27 SOLiD, one Ion Torrent PGM, one MiSeq, and one 454 sequencer. We have accumulated extensive experience in sample handling, sequencing, and bioinformatics analysis. In this paper, technologies of these systems are reviewed, and first-hand data from extensive experience is summarized and analyzed to discuss the advantages and specifics associated with each sequencing system. At last, applications of NGS are summarized.

  4. Analysis of autocorrelation and ambiguity function of complicated radiolocation signal with binary phase manipulation by Kasami sequence

    Directory of Open Access Journals (Sweden)

    O. D. Mrachkovskyi

    2015-09-01

    Full Text Available Introduction. Were reviewed publications dedicated to pseudorandom sequences in radiolocation, and was made conclusion about necessity of researching in this subject. Modeling of probing radiolocation signal with binary phase manipulation by Kasami sequence. Were considered the method of forming Kasami sequences and measuring their properties. Example of generator for sequence with length 63 symbols is also shown. Technique of research. Were shown graphical results for modeling of Kasami sequence with lengths of 255 binary values. Amplitude of side maximums of autocorrelation function are measured. Modeling of ambiguity function. In this article, graphical results for modeling Kasami sequence ambiguity function are shown. These includes 3-dimensional plot of function body and its cross-sections by plane of equal ranges and planes of function level, equal to 0.9, 0.7, 0.5, 0.4, 0.3, 0.2 and 0.1 of its maximum. Were analyzed amplitude of side maximums, cross-section form and relative values of range and speed resolution. Conclusion. These articles contain analyses of characteristics of previously shown plots. These analyses include estimation of total value of side maximums of ambiguity function of Kasami sequence, ambiguity function cross-section form analysis and conclusion about ability of using these sequences as probe signal in radiolocation.

  5. White dwarf-main sequence binaries from LAMOST: the DR1 catalogue

    CERN Document Server

    Ren, Juanjuan; Luo, Ali; Zhao, Yongheng; Xiang, Maosheng; Liu, Xiaowei; Zhao, Gang; Jin, Ge; Zhang, Yong

    2014-01-01

    Context. White dwarf-main sequence (WDMS) binaries are used to study several different important open problems in modern astrophysics. Aims. The Sloan Digital Sky Survey (SDSS) identified the largest catalogue of WDMS binaries currently known. However, this sample is seriously affected by selection effects and the population of systems containing cool white dwarfs and early-type companions is under-represented.Here we search for WDMS binaries within the spectroscopic data release 1 of the LAMOST (Large sky Area Multi-Object fiber Spectroscopic Telescope) survey. LAMOST and SDSS follow different target selection algorithms. Hence, LAMOST WDMS binaries may be drawn from a different parent population and thus help in overcoming the selection effects incorporated by SDSS on the current observed population. Methods. We develop a fast and efficient routine based on the wavelet transform to identify LAMOST WDMS binaries containing a DA white dwarf and a M dwarf companion, and apply a decomposition/fitting routine to...

  6. Detecting black-hole binary clustering via the second-generation gravitational-wave detectors

    OpenAIRE

    Namikawa, Toshiya; Nishizawa, Atsushi; Taruya, Atsushi

    2016-01-01

    The first discovery of the gravitational wave (GW) event, GW150914, suggests a higher merger rate of black-hole (BH) binaries. If this is true, a number of BH binaries will be observed via the second-generation GW detectors, and the statistical properties of the observed BH binaries can be scrutinized. A naive but important question to ask is whether the spatial distribution of BH binaries faithfully traces the matter inhomogeneities in the Universe or not. Although the BH binaries are though...

  7. Orbital Parameters for a Pre-Main Sequence Binary System

    Science.gov (United States)

    Karnath, Nicole; Prato, L.; Wasserman, L.

    2011-01-01

    The young system VSB 111 was originally classified as a single-lined spectroscopic binary in the star forming region of NGC 2264. Using the Keck II telescope we measured radial velocities for both the primary and secondary components in the infrared. By combining these data with previous visible light observations of the primary star, we derived the period, eccentricity, and other orbital parameters, as well as the mass ratio of the system. With additional information gained from further observations, for example the inclination derived from the angularly resolved orbit, we will eventually obtain the individual stellar masses, necessary to help to calibrate models of young star evolution. Furthermore, by compiling dozens or even hundreds of mass ratios for young binaries we can use mass ratio distributions to improve our understanding of binary star formation. No infrared excess or any other indication of a circumstellar disk is in evidence for VSB 111, indicating that either the accretion rate has dropped to an undetectable value or that this system has aged enough that its disk has dissipated, if originally present. Given the approximately 900 day period of this system, and its relatively high eccentricity, 0.8, the action of the companion could have been responsible for early dissipation of any disk material.

  8. Main-Sequence Binary Stars in the Core of NGC 6397

    Science.gov (United States)

    Bolton, A. S.; Cool, A. M.; Anderson, J.

    1999-12-01

    Using HST WFPC2 data, we isolate main-sequence binary candidates in the central region of the globular cluster NGC 6397 based on their locations in an I vs. V - I color--magnitude diagram. We have largely eliminated field stars from the sample beforehand based on proper motions determined from two sets of position data separated by approximately three years. Binary candidates are fit to models based on the empirically derived main-sequence ridge line for the cluster, and component masses are determined using theoretical mass--luminosity relations appropriate to the cluster. Preliminary results suggest an upper limit of 3% on the binary fraction for stars in the apparent magnitude range 17.0 mass ratios greater than approximately 0.45. We also present preliminary results for the distribution of binaries as a function of primary mass and mass ratio, as well as a comparison of these results to previously published findings for field stars.

  9. Discovery of ZZ Cetis in detached white dwarf plus main-sequence binaries

    CERN Document Server

    Pyrzas, S; Hermes, J J; Copperwheat, C M; Rebassa-Mansergas, A; Dhillon, V S; Littlefair, S P; Marsh, T R; Parsons, S G; Savoury, C D J; Schreiber, M R; Barros, S C C; Bento, J; Breedt, E; Kerry, P

    2014-01-01

    We present the first results of a dedicated search for pulsating white dwarfs (WDs) in detached white dwarf plus main-sequence binaries. Candidate systems were selected from a catalogue of WD+MS binaries, based on the surface gravities and effective temperatures of the WDs. We observed a total of 26 systems using ULTRACAM mounted on ESO's 3.5m New Technology Telescope (NTT) at La Silla. Our photometric observations reveal pulsations in seven WDs of our sample, including the first pulsating white dwarf with a main-sequence companion in a post common envelope binary, SDSSJ1136+0409. Asteroseismology of these new pulsating systems will provide crucial insight into how binary interactions, particularly the common envelope phase, affect the internal structure and evolution of WDs. In addition, our observations have revealed the partially eclipsing nature of one of our targets, SDSSJ1223-0056.

  10. A binary sequence of period 60 with better autocorrelation properties than the Barker sequence of period 13

    Science.gov (United States)

    Watkins, J.; Loftsson, J.; Tyler, S.

    1985-01-01

    A binary sequence of period 60 has been discovered which in some respects has better autocorrelation properties than the Barker sequence of period 13. When both sequences are processed using appropriate sidelobe-eliminating mismatched filters, the Barker sequence's main lobe is reduced by a factor of 1.040 or 0.17 dB, while the new sequence's main lobe is reduced by a factor of only 1.035 or 0.15 dB. This sequence is the first counterexample known to the authors of the hypothesis that the autocorrelation properties of all sequences of periods greater than 13 are inferior to those of the Barker period-13 sequences. Sequences of this type are very useful in radar and deep space communications, especially in situations where there is an adverse signal to noise ratio.

  11. Exploiting mid-range DNA patterns for sequence classification: binary abstraction Markov models

    Science.gov (United States)

    Shepard, Samuel S.; McSweeny, Andrew; Serpen, Gursel; Fedorov, Alexei

    2012-01-01

    Messenger RNA sequences possess specific nucleotide patterns distinguishing them from non-coding genomic sequences. In this study, we explore the utilization of modified Markov models to analyze sequences up to 44 bp, far beyond the 8-bp limit of conventional Markov models, for exon/intron discrimination. In order to analyze nucleotide sequences of this length, their information content is first reduced by conversion into shorter binary patterns via the application of numerous abstraction schemes. After the conversion of genomic sequences to binary strings, homogenous Markov models trained on the binary sequences are used to discriminate between exons and introns. We term this approach the Binary Abstraction Markov Model (BAMM). High-quality abstraction schemes for exon/intron discrimination are selected using optimization algorithms on supercomputers. The best MM classifiers are then combined using support vector machines into a single classifier. With this approach, over 95% classification accuracy is achieved without taking reading frame into account. With further development, the BAMM approach can be applied to sequences lacking the genetic code such as ncRNAs and 5′-untranslated regions. PMID:22344692

  12. Galaxy LIMS for next-generation sequencing

    NARCIS (Netherlands)

    Scholtalbers, J.; Rossler, J.; Sorn, P.; Graaf, J. de; Boisguerin, V.; Castle, J.; Sahin, U.

    2013-01-01

    SUMMARY: We have developed a laboratory information management system (LIMS) for a next-generation sequencing (NGS) laboratory within the existing Galaxy platform. The system provides lab technicians standard and customizable sample information forms, barcoded submission forms, tracking of input sam

  13. Fetal Kidney Anomalies: Next Generation Sequencing

    DEFF Research Database (Denmark)

    Rasmussen, Maria; Sunde, Lone; Nielsen, Marlene Louise

    Aim and Introduction Identification of abnormal kidneys in the fetus may lead to termination of the pregnancy and raises questions about the underlying cause and recurrence risk in future pregnancies. In this study, we investigate the effectiveness of targeted next generation sequencing in fetuses...

  14. ANGSD: Analysis of Next Generation Sequencing Data.

    Science.gov (United States)

    Korneliussen, Thorfinn Sand; Albrechtsen, Anders; Nielsen, Rasmus

    2014-11-25

    High-throughput DNA sequencing technologies are generating vast amounts of data. Fast, flexible and memory efficient implementations are needed in order to facilitate analyses of thousands of samples simultaneously. We present a multithreaded program suite called ANGSD. This program can calculate various summary statistics, and perform association mapping and population genetic analyses utilizing the full information in next generation sequencing data by working directly on the raw sequencing data or by using genotype likelihoods. The open source c/c++ program ANGSD is available at http://www.popgen.dk/angsd . The program is tested and validated on GNU/Linux systems. The program facilitates multiple input formats including BAM and imputed beagle genotype probability files. The program allow the user to choose between combinations of existing methods and can perform analysis that is not implemented elsewhere.

  15. RAMBO-K: Rapid and Sensitive Removal of Background Sequences from Next Generation Sequencing Data.

    Directory of Open Access Journals (Sweden)

    Simon H Tausch

    Full Text Available The assembly of viral or endosymbiont genomes from Next Generation Sequencing (NGS data is often hampered by the predominant abundance of reads originating from the host organism. These reads increase the memory and CPU time usage of the assembler and can lead to misassemblies.We developed RAMBO-K (Read Assignment Method Based On K-mers, a tool which allows rapid and sensitive removal of unwanted host sequences from NGS datasets. Reaching a speed of 10 Megabases/s on 4 CPU cores and a standard hard drive, RAMBO-K is faster than any tool we tested, while showing a consistently high sensitivity and specificity across different datasets.RAMBO-K rapidly and reliably separates reads from different species without data preprocessing. It is suitable as a straightforward standard solution for workflows dealing with mixed datasets. Binaries and source code (java and python are available from http://sourceforge.net/projects/rambok/.

  16. Phase retrieval from multiple binary masks generated speckle patterns

    Science.gov (United States)

    Gong, Hai; Pozzi, Paolo; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

    2016-04-01

    We present a reference-less and time-multiplexing phase retrieval method by making use of the digital micromirror device (DMD). In this method, the DMD functions not only as a flexible binary mask which modulates the optical field, but also as a sampling mask for measuring corresponding phases, which makes the whole setup simple and robust. The DMD reflection forms a sparse intensity mask in the pupil which produces speckle pattern after propagation. With the recorded intensity on the camera and the binary pattern on the DMD, the phase in all the `on' pixels can be reconstructed at once by solving inverse problems with iterative methods, for instance using Gerchberg-Saxton algorithm. Then the phase of the whole pupil can be reconstructed from a series of binary patterns and speckle patterns. Numerical experiments show the feasibility of this phase retrieval method and the importance of sparse binary masks in the improving of convergence speed.

  17. Generation of artificial FASTQ files to evaluate the performance of next-generation sequencing pipelines.

    Directory of Open Access Journals (Sweden)

    Matthew Frampton

    Full Text Available Pipelines for the analysis of Next-Generation Sequencing (NGS data are generally composed of a set of different publicly available software, configured together in order to map short reads of a genome and call variants. The fidelity of pipelines is variable. We have developed ArtificialFastqGenerator, which takes a reference genome sequence as input and outputs artificial paired-end FASTQ files containing Phred quality scores. Since these artificial FASTQs are derived from the reference genome, it provides a gold-standard for read-alignment and variant-calling, thereby enabling the performance of any NGS pipeline to be evaluated. The user can customise DNA template/read length, the modelling of coverage based on GC content, whether to use real Phred base quality scores taken from existing FASTQ files, and whether to simulate sequencing errors. Detailed coverage and error summary statistics are outputted. Here we describe ArtificialFastqGenerator and illustrate its implementation in evaluating a typical bespoke NGS analysis pipeline under different experimental conditions. ArtificialFastqGenerator was released in January 2012. Source code, example files and binaries are freely available under the terms of the GNU General Public License v3.0. from https://sourceforge.net/projects/artfastqgen/.

  18. Tablet—next generation sequence assembly visualization

    OpenAIRE

    Milne, Iain; Bayer, Micha; Cardle, Linda; Shaw, Paul; Stephen, Gordon; Wright, Frank; Marshall, David

    2009-01-01

    Summary: Tablet is a lightweight, high-performance graphical viewer for next-generation sequence assemblies and alignments. Supporting a range of input assembly formats, Tablet provides high-quality visualizations showing data in packed or stacked views, allowing instant access and navigation to any region of interest, and whole contig overviews and data summaries. Tablet is both multi-core aware and memory efficient, allowing it to handle assemblies containing millions of reads, even on a 32...

  19. Stochastic Background of Gravitational Waves Generated by Compact Binary Systems

    CERN Document Server

    Evangelista, E F D

    2015-01-01

    Binary Systems are the most studied sources of gravitational waves. The mechanisms of emission and the behavior of the orbital parameters are well known and can be written in analytic form in several cases. Besides, the strongest indication of the existence of gravitational waves has arisen from the observation of binary systems. On the other hand, when the detection of gravitational radiation becomes a reality, one of the observed pattern of the signals will be probably of stochastic background nature, which are characterized by a superposition of signals emitted by many sources around the universe. Our aim here is to develop an alternative method of calculating such backgrounds emitted by cosmological compact binary systems during their periodic or quasiperiodic phases. We use an analogy with a problem of Statistical Mechanics in order to perform this sum as well as taking into account the temporal variation of the orbital parameters of the systems. Such a kind of background is of particular importance sinc...

  20. Binary star influence on post-main-sequence multi-planet stability

    CERN Document Server

    Veras, Dimitri; Dobbs-Dixon, Ian; Gaensicke, Boris T

    2016-01-01

    Nearly every star known to host planets will become a white dwarf, and nearly 100 planet-hosts are now known to be accompanied by binary stellar companions. Here, we determine how a binary companion triggers instability in otherwise unconditionally stable single-star two-planet systems during the giant branch and white dwarf phases of the planet host. We perform about 700 full-lifetime (14 Gyr) simulations with A0 and F0 primary stars and secondary K2 companions, and identify the critical binary distance within which instability is triggered at any point during stellar evolution. We estimate this distance to be about seven times the outer planet separation, for circular binaries. Our results help characterize the fates of planetary systems, and in particular which ones might yield architectures that are conducive to generating observable heavy metal pollution in white dwarf atmospheres.

  1. Binary star influence on post-main-sequence multi-planet stability

    Science.gov (United States)

    Veras, Dimitri; Georgakarakos, Nikolaos; Dobbs-Dixon, Ian; Gänsicke, Boris T.

    2017-02-01

    Nearly every star known to host planets will become a white dwarf, and nearly 100 planet-hosts are now known to be accompanied by binary stellar companions. Here, we determine how a binary companion triggers instability in otherwise unconditionally stable single-star two-planet systems during the giant branch and white dwarf phases of the planet host. We perform about 700 full-lifetime (14 Gyr) simulations with A0 and F0 primary stars and secondary K2 companions, and identify the critical binary distance within which instability is triggered at any point during stellar evolution. We estimate this distance to be about seven times the outer planet separation for circular binaries. Our results help characterize the fates of planetary systems, and in particular which ones might yield architectures which are conducive to generating observable metal pollution in white dwarf atmospheres.

  2. Differential rotation on both components of the pre main-sequence binary system HD 155555

    OpenAIRE

    Dunstone, N. J.; Hussain, G A J; Cameron, A. Collier; Marsden, S. C.; Jardine, M.; Barnes, J. R.; Vlex, J. C. Ramirez; Donati, J.-F.

    2008-01-01

    We present the first measurements of surface differential rotation on a pre-main sequence binary system. Using intensity (Stokes I) and circularly polarised (Stokes V) timeseries spectra, taken over eleven nights at the Anglo-Australian Telescope (AAT), we incorporate a solar-like differential rotation law into the surface imaging process. We find that both components of the young, 18 Myr, HD 155555 (V824 Ara, G5IV + K0IV) binary system show significant differential rotation. The equator-pole...

  3. Quasi-equilibrium sequences of binary strange quark stars in general relativity

    Science.gov (United States)

    Limousin, Francois; Gondek-Rosińska, Dorota; Gourgoulhon, Eric

    2004-12-01

    Inspiraling compact binaries are expected to be the strongest sources of gravitational waves for VIRGO, LIGO and other laser interferometers. We present the first computations of quasi-equilibrium sequences of compact binaries containing two strange quark stars (which are currently considered as a possible alternative to neutron stars). We study a precoalescing stage in the conformal flatness approximation of general relativity using a multidomain spectral method. A hydrodynamical treatment is performed under the assumption that the flow is either rigidly rotating or irrotational. In each of those cases, we show the differences in the gravitational waves signal from neutron stars described by polytropic equation of state.

  4. The first magnetic maps of a pre-main sequence binary star system - HD 155555

    OpenAIRE

    Dunstone, N. J.; Hussain, G. A. J.; Cameron, A. Collier; Marsden, S. C.; Jardine, M.; Stempels, H. C.; Vlex, J. C. Ramirez; Donati, J. -F.

    2008-01-01

    We present the first maps of the surface magnetic fields of a pre-main sequence binary system. Spectropolarimetric observations of the young, 18 Myr, HD 155555 (V824 Ara, G5IV + K0IV) system were obtained at the Anglo-Australian Telescope in 2004 and 2007. Both datasets are analysed using a new binary Zeeman Doppler imaging (ZDI) code. This allows us to simultaneously model the contribution of each component to the observed circularly polarised spectra. Stellar brightness maps are also produc...

  5. Differential rotation on both components of the pre main-sequence binary system HD 155555

    OpenAIRE

    Dunstone, N. J.; Hussain, G. A. J.; Cameron, A. Collier; Marsden, S. C.; Jardine, M.; Barnes, J.R.; Vlex, J. C. Ramirez; Donati, J. -F.

    2008-01-01

    We present the first measurements of surface differential rotation on a pre-main sequence binary system. Using intensity (Stokes I) and circularly polarised (Stokes V) timeseries spectra, taken over eleven nights at the Anglo-Australian Telescope (AAT), we incorporate a solar-like differential rotation law into the surface imaging process. We find that both components of the young, 18 Myr, HD 155555 (V824 Ara, G5IV + K0IV) binary system show significant differential rotation. The equator-pole...

  6. Generating matrix and sums of Fibonacci and Pell sequences

    Science.gov (United States)

    Ho, C. K.; Woon, H. S.; Chong, Chin-Yoon

    2014-07-01

    In this paper, we study the Fibonacci sequence and Pell sequence and developed generating matrices for them. First we proved two results on the even sum of the Fibonacci sequence and the Pell sequence, using the generating matrix approach. We then deduce the odd sums, some identities and recursive formulas for these two sequences.

  7. Detecting black-hole binary clustering via the second-generation gravitational-wave detectors

    Science.gov (United States)

    Namikawa, Toshiya; Nishizawa, Atsushi; Taruya, Atsushi

    2016-07-01

    The first discovery of the gravitational-wave (GW) event, GW150914, suggests a higher merger rate of black-hole (BH) binaries. If this is true, a number of BH binaries will be observed via the second-generation GW detectors, and the statistical properties of the observed BH binaries can be scrutinized. A naive but important question to ask is whether the spatial distribution of BH binaries faithfully traces the matter inhomogeneities in the Universe or not. Although the BH binaries are thought to be formed inside the galaxies in most of the scenarios, there is no observational evidence to confirm such a hypothesis. Here, we estimate how well the second-generation GW detectors can statistically confirm the BH binaries to be a tracer of the large-scale structure by looking at the auto- and cross-correlation of BH binaries with photometric galaxies and weak-lensing measurements, finding that, with a 3 year observation, the >3 σ detection of a nonzero signal is possible if the BH merger rate today is n˙ 0≳100 Gpc-3 yr-1 and the clustering bias of BH binaries is bBH ,0≳1.5 .

  8. Statistical analysis of next generation sequencing data

    CERN Document Server

    Nettleton, Dan

    2014-01-01

    Next Generation Sequencing (NGS) is the latest high throughput technology to revolutionize genomic research. NGS generates massive genomic datasets that play a key role in the big data phenomenon that surrounds us today. To extract signals from high-dimensional NGS data and make valid statistical inferences and predictions, novel data analytic and statistical techniques are needed. This book contains 20 chapters written by prominent statisticians working with NGS data. The topics range from basic preprocessing and analysis with NGS data to more complex genomic applications such as copy number variation and isoform expression detection. Research statisticians who want to learn about this growing and exciting area will find this book useful. In addition, many chapters from this book could be included in graduate-level classes in statistical bioinformatics for training future biostatisticians who will be expected to deal with genomic data in basic biomedical research, genomic clinical trials and personalized med...

  9. KH 15D: Gradual Occultation of a Pre-Main-Sequence Binary

    CERN Document Server

    Winn, J N; Johnson, J A; Stanek, K Z; Garnavich, P M; Winn, Joshua N.; Holman, Matthew J.; Johnson, John A.; Stanek, Krzysztof Z.; Garnavich, Peter M.

    2004-01-01

    We propose that the extraordinary "winking star" KH 15D is an eccentric pre-main-sequence binary that is gradually being occulted by an opaque screen. This model accounts for the periodicity, depth, duration, and rate of growth of the modern eclipses; the historical light curve from photographic plates; and the existing radial velocity measurements. It also explains the re-brightening events that were previously observed during mid-eclipse, and the subsequent disappearance of these events. We predict the future evolution of the system and its full radial velocity curve. Given the small velocity of the occulting screen relative to the center of mass of the binary, the screen is probably associated with the binary, and may be the edge of a precessing circumbinary disk.

  10. Detecting Black-Hole Binary Clustering via the Second-Generation Gravitational-Wave Detectors

    CERN Document Server

    Namikawa, Toshiya; Taruya, Atsushi

    2016-01-01

    First discovery of the gravitational wave (GW) event, GW150914, suggests a higher merger rate of black-hole (BH) binaries. If this is true, a number of BH binaries will be observed via the second-generation GW detectors, and the statistical properties of the observed BH binaries can be scrutinized. A naive but important question to ask is whether the spatial distribution of BH binaries faithfully traces the matter inhomogeneities in the Universe or not. Although the BH binaries are thought to be formed inside the galaxies in most of the scenarios, there is no observational evidence to confirm such a hypothesis. Here, we estimate how well the second-generation GW detectors can statistically confirm the BH binaries to be a tracer of the large-scale structure by looking at the auto- and cross-correlation of BH binaries with photometric galaxies and weak lensing measurements, finding that, with a three-year observation, the $>3\\sigma$ detection of non-zero signal is possible if the BH merger rate today is $\\dot{n...

  11. Period of the d-Sequence Based Random Number Generator

    OpenAIRE

    Thippireddy, Suresh; Chalasani, Sandeep

    2007-01-01

    This paper presents an expression to compute the exact period of a recursive random number generator based on d-sequences. Using the multi-recursive version of this generator we can produce large number of pseudorandom sequences.

  12. Compact object detection in self-lensing binary systems with a main-sequence star

    CERN Document Server

    Rahvar, S; Dominik, M

    2010-01-01

    Detecting compact objects by means of their gravitational lensing effect on an observed companion in a binary system has already been suggested almost four decades ago. However, these predictions were made even before the first observations of gravitational lensing, whereas nowadays gravitational microlensing surveys towards the Galactic bulge yield almost 1000 events per year where one star magnifies the light of a more distant one. With a specific view on those experiments, we therefore carry out simulations to assess the prospects for detection of the transient periodic magnification of the companion star, which lasts typically only a few hours binaries involving a main-sequence star. We find that detectability is given by the achievability of dense monitoring with the required photometric accuracy. In sharp contrast to earlier expectations by other authors, we find that main-sequence stars are not substantially less favourable targets to observe this effect than white dwarfs. The requirement of an almost ...

  13. Hunting for millimeter flares from magnetic reconnection in pre-main sequence spectroscopic binaries

    CERN Document Server

    Kóspál, Á; Hogerheijde, M R; Moór, A; Blake, G A

    2010-01-01

    Recent observations of the low-mass pre-main sequence, eccentric spectroscopic binaries DQ Tau and V773 Tau A reveal that their millimeter spectrum is occasionally dominated by flares from non-thermal emission processes. The transient activity is believed to be synchrotron in nature, resulting from powerful magnetic reconnection events when the separate magnetic structures of the binary components are capable of interacting and forced to reorganize, typically near periastron. We conducted the first systematic study of the millimeter variability toward a sample of 12 PMS spectroscopic binaries with the aim to characterize the proliferation of flares amongst sources likely to experience similar interbinary reconnection events. The source sample consists of short-period, close-separation binaries that possess either a high orbital eccentricity or a circular orbit. Using the MAMBO2 array on the IRAM 30m telescope, we carried out continuous monitoring at 1.25 mm over a 4-night period during which all of the high-e...

  14. The population of white dwarf-main sequence binaries in the SDSS DR 12

    Science.gov (United States)

    Cojocaru, R.; Rebassa-Mansergas, A.; Torres, S.; García-Berro, E.

    2017-09-01

    We present a Monte Carlo population synthesis study of white dwarf-main sequence (WD+MS) binaries in the Galactic disc aimed at reproducing the ensemble properties of the entire population observed by the Sloan Digital Sky Survey (SDSS) Data Release 12. Our simulations take into account all known observational biases and use the most up-to-date stellar evolutionary models. This allows us to perform a sound comparison between the simulations and the observational data. We find that the properties of the simulated and observed parameter distributions agree best when assuming low values of the common envelope efficiency (0.2-0.3), a result that is in agreement with previous findings obtained by observational and population synthesis studies of close SDSS WD+MS binaries. We also show that all synthetic populations that result from adopting an initial mass ratio distribution with a positive slope are excluded by observations. Finally, we confirm that the properties of the simulated WD+MS binary populations are nearly independent of the age adopted for the thin disc, on the contribution of WD+MS binaries from the thick disc (0-17 per cent of the total population) and on the assumed fraction of the internal energy that is used to eject the envelope during the common envelope phase (0.1-0.5).

  15. Application of next-generation sequencing for comparative transcriptome analysis

    OpenAIRE

    Shin, Heesun

    2010-01-01

    I have used novel whole transcriptome sequence data generated from massively parallel high-throughput next generation sequencing technologies, namely 454 pyrosequencing and Illumina sequencing, to perform comparative transcriptome analyses of C. elegans populations in specific biological conditions and developmental stages. Firstly, I have conducted transcriptome profiling of C. elegans in its first larval (L1) stage using data generated from the Roche 454 sequencing platform. I have used thi...

  16. Preparation of SELEX Samples for Next-Generation Sequencing.

    Science.gov (United States)

    Tolle, Fabian; Mayer, Günter

    2016-01-01

    Fuelled by massive whole genome sequencing projects such as the human genome project, enormous technological advancements and therefore tremendous price drops could be achieved, rendering next-generation sequencing very attractive for deep sequencing of SELEX libraries. Herein we describe the preparation of SELEX samples for Illumina sequencing, based on the already established whole genome sequencing workflow. We describe the addition of barcode sequences for multiplexing and the adapter ligation, avoiding associated pitfalls.

  17. New theoretical bounds on the aperiodic correlation functions of binary sequences

    Institute of Scientific and Technical Information of China (English)

    PENG Daiyuan; FAN Pingzhi

    2005-01-01

    In order to reduce or eliminate the multiple access interference in code division multiple access (CDMA) systems, we need to design a set of spreading sequences with good autocorrelation functions (ACF) and crosscorrelation functions (CCF). The importance of the spreading codes to CDMA systems cannot be overemphasized, for the type of the code used, its length, and its chip rate set bounds on the capability of the system that can be changed only by changing the code. Several new lower bounds which are stronger than the well-known Sarwate bounds, Welch bounds and Levenshtein bounds for binary sequence set with respect to the spreading sequence length, family size, maximum aperiodic autocorrelation sidelobe and maximum aperiodic crosscorrelation value are established.

  18. Spectrum analysis of complicated radiolocation signal with binary phase manipulation by Kasami sequence

    Directory of Open Access Journals (Sweden)

    O. D. Mrachkovskyi

    2015-06-01

    Full Text Available Introduction. This article contain short introduction to history of complicated signals and list of common sequences, used in spectrum spreading. Method of creating Kasami sequences. In this article is shown sequence creation method, based on method of creating maximum length sequence. Creating of the structural schema of Kasami sequences generator. Principles of construction of Kasami sequence generator are shown in this article. Example of generator for sequence with length 63 symbols, which are created using method, mentioned in previous article, is also shown. Spectrum of Kasami sequences. This article contain amplitude and phase spectrum of two Kasami sequences with lengths of 1023 and 255 symbols. Theoretical formulas are created using Discrete Fourier Transform operation, and graphics are plotted using Fast Fourier Transform operation. Phase spectrum data are also been changed by unwrap operation for better visibility. Conclusion. This article contain analysis of characteristics of previously shown spectrum. Several conclusions are made about correlation between sequence and spectrum characteristics. These conclusions include that made about amplitude spectrum form dependence of sequence length, type and basic impulse characteristics.

  19. Generating Functions for the Powers of Fibonacci Sequences

    Science.gov (United States)

    Terrana, D.; Chen, H.

    2007-01-01

    In this note, based on the Binet formulas and the power-reducing techniques, closed forms of generating functions for the powers of Fibonacci sequences are presented. The corresponding results are extended to some other famous sequences as well.

  20. Next-generation sequencing offers new insights into DNA degradation

    DEFF Research Database (Denmark)

    Overballe-Petersen, Søren; Orlando, Ludovic Antoine Alexandre; Willerslev, Eske

    2012-01-01

    The processes underlying DNA degradation are central to various disciplines, including cancer research, forensics and archaeology. The sequencing of ancient DNA molecules on next-generation sequencing platforms provides direct measurements of cytosine deamination, depurination and fragmentation r...

  1. Computer-Generated Phase Diagrams for Binary Mixtures.

    Science.gov (United States)

    Jolls, Kenneth R.; And Others

    1983-01-01

    Computer programs that generate projections of thermodynamic phase surfaces through computer graphics were used to produce diagrams representing properties of water and steam and the pressure-volume-temperature behavior of most of the common equations of state. The program, program options emphasizing thermodynamic features of interest, and…

  2. The k-Error Linear Complexity and the Linear Complexity for pqn-Periodic Binary Sequences

    Institute of Scientific and Technical Information of China (English)

    ZHU Fengxiang; QI Wenfeng

    2006-01-01

    The k-error linear complexity and the linear complexity of the keystream of a stream cipher are two important standards to scale the randomness of the key stream. For a pqn-periodic binary sequences where p, q are two odd primes satisfying that 2 is a primitive root module p and q2 and gcd(p-1 , q-1)=2, we analyze the relationship between the linear complexity and the minimum value k for which the k-error linear complexity is strictly less than the linear complexity.

  3. Generating barcoded libraries for multiplex high-throughput sequencing.

    Science.gov (United States)

    Knapp, Michael; Stiller, Mathias; Meyer, Matthias

    2012-01-01

    Molecular barcoding is an essential tool to use the high throughput of next generation sequencing platforms optimally in studies involving more than one sample. Various barcoding strategies allow for the incorporation of short recognition sequences (barcodes) into sequencing libraries, either by ligation or polymerase chain reaction (PCR). Here, we present two approaches optimized for generating barcoded sequencing libraries from low copy number extracts and amplification products typical of ancient DNA studies.

  4. Metagenomics using next-generation sequencing.

    Science.gov (United States)

    Bragg, Lauren; Tyson, Gene W

    2014-01-01

    Traditionally, microbial genome sequencing has been restricted to the small number of species that can be grown in pure culture. The progressive development of culture-independent methods over the last 15 years now allows researchers to sequence microbial communities directly from environmental samples. This approach is commonly referred to as "metagenomics" or "community genomics". However, the term metagenomics is applied liberally in the literature to describe any culture-independent analysis of microbial communities. Here, we define metagenomics as shotgun ("random") sequencing of the genomic DNA of a sample taken directly from the environment. The metagenome can be thought of as a sampling of the collective genome of the microbial community. We outline the considerations and analyses that should be undertaken to ensure the success of a metagenomic sequencing project, including the choice of sequencing platform and methods for assembly, binning, annotation, and comparative analysis.

  5. Optical image encryption based on binary Fourier transform computer-generated hologram and pixel scrambling technology

    Science.gov (United States)

    Wang, Yong-Ying; Wang, Yu-Rong; Wang, Yong; Li, Hui-Juan; Sun, Wen-Jia

    2007-07-01

    A new method of optical image encryption with binary Fourier transform computer-generated hologram (CGH) and pixel-scrambling technology is presented. In this method, the orders of the pixel scrambling, as well as the encrypted image, are used as the keys to decrypt the original image. Therefore, higher security is achieved. Furthermore, the encrypted image is binary, so it is easy to be fabricated and robust against noise and distortion. Computer simulation results are given to verify the feasibility of this method and its robustness against occlusion and additional noise.

  6. Differential rotation on both components of the pre main-sequence binary system HD 155555

    CERN Document Server

    Dunstone, N J; Cameron, A Collier; Marsden, S C; Jardine, M; Barnes, J R; Vlex, J C Ramirez; Donati, J -F

    2008-01-01

    We present the first measurements of surface differential rotation on a pre-main sequence binary system. Using intensity (Stokes I) and circularly polarised (Stokes V) timeseries spectra, taken over eleven nights at the Anglo-Australian Telescope (AAT), we incorporate a solar-like differential rotation law into the surface imaging process. We find that both components of the young, 18 Myr, HD 155555 (V824 Ara, G5IV + K0IV) binary system show significant differential rotation. The equator-pole laptimes as determined from the intensity spectra are 80 days for the primary star and 163 days for the secondary. Similarly for the magnetic spectra we obtain equator-pole laptimes of 44 and 71 days respectively, showing that the shearing timescale of magnetic regions is approximately half that found for stellar spots. Both components are therefore found to have rates of differential rotation similar to those of the same spectral type main sequence single stars. The results for HD 155555 are therefore in contrast to tho...

  7. Evidence of accretion triggered oscillations in the pre-main-sequence interacting binary AK Sco

    CERN Document Server

    de Castro, Ana I Gomez; Talavera, Antonio

    2012-01-01

    Pre-main sequence (PMS) binaries are surrounded by circumbinary disks from which matter falls onto both components. The material dragged from the circumbinary disk flows onto each star through independent streams channelled by the variable gravitational field. The action of the bar-like potential is most prominent in high eccentricity systems made of two equal mass stars. AK Sco is a unique PMS system composed of two F5 stars in an orbit with e=0.47. Henceforth, it is an ideal laboratory to study matter infall in binaries and its role in orbit circularization. In this letter, we report the detection of a 1.3mHz ultra low frequency oscillation in the ultraviolet light curve at periastron passage. This oscillation last 7 ks being most likely fed by the gravitational energy released when the streams tails spiralling onto each star get in contact at periastron passage enhancing the accretion flow; this unveils a new mechanism for angular momentum loss during pre-main sequence evolution and a new type of interacti...

  8. A LARGE CLASS OF BINARY ZCZ SEQUENCE FAMILIES CONSTRUCTED BY PERIOD DOUBLING

    Institute of Scientific and Technical Information of China (English)

    Wang Jinsong; Qi Wenfeng

    2007-01-01

    In an Approximately Synchronized Code Division Multiple Access(AS-CDMA)communication svstem,a family with large number of Zero Correlation Zone(ZCZ)sequences is desired,which can satisfy the rapid increase of users.This paper presents a method to generate a(2L,2M,Z'cz)-ZCZ sequence family from an original(L,M,Zcz)-ZCZ sequence family,where Z'cz=Zcz if Zcz is even and Z'cz=Zcz-1 if Zcz is odd.This method can also recursively act on a ZCZ sequence family to construct a series of ZCZ sequence families with large sequence nutuber and zero correlation zone length identical to or one less than that of original ZCZ sequences.

  9. Chaotic block iterating method for pseudo-random sequence generator

    Institute of Scientific and Technical Information of China (English)

    CHEN Shuai; ZHONG Xian-xin

    2007-01-01

    A pseudo-random sequence generator is a basic tool for cryptography. To realize a pseudo-random sequence generator, a new block iterating method using shifter, multiplier,and adder operations has been introduced. By increasing the iteration of the counter and by performing calculations based on the initial value, an approximate pseudo-random sequence was obtained after exchanging bits. The algorithm and the complexity of the generator were introduced. The result obtained from the calculation shows that the self-correlation of the "m" block sequence is two-valued; the block field value is [0,2m - 1 ], and the block period is 2m+8 - 1.

  10. Pseudo-Random Sequences Generator Based on Discrete Hyperchaotic Systems

    Institute of Scientific and Technical Information of China (English)

    李昌刚; 韩正之

    2003-01-01

    We first design a discrete hyperchaotic system via piecewise linear state feedback. The states of the closed loop system are locally expanding in two directions but absolutely bounded on the whole, which implies hyperchaos. Then, we use three suchlike hyperchaotie systems with different feedback gain matrices to design a pseudo-random sequence generator (PRSG). Through a threshold function, three sub-sequences generated from the output of piecewise linear functions are changed into 0-1 sequences. Then, followed by XOR operation, an unpredictable pseudo-random sequence (PRS) is ultimately obtained. The analysis and simulation results indicate that the PRS, generated with hyperchaotic systems, has desirable statistical features.

  11. Image encryption using random sequence generated from generalized information domain

    Science.gov (United States)

    Xia-Yan, Zhang; Guo-Ji, Zhang; Xuan, Li; Ya-Zhou, Ren; Jie-Hua, Wu

    2016-05-01

    A novel image encryption method based on the random sequence generated from the generalized information domain and permutation-diffusion architecture is proposed. The random sequence is generated by reconstruction from the generalized information file and discrete trajectory extraction from the data stream. The trajectory address sequence is used to generate a P-box to shuffle the plain image while random sequences are treated as keystreams. A new factor called drift factor is employed to accelerate and enhance the performance of the random sequence generator. An initial value is introduced to make the encryption method an approximately one-time pad. Experimental results show that the random sequences pass the NIST statistical test with a high ratio and extensive analysis demonstrates that the new encryption scheme has superior security.

  12. Automated Testing with Targeted Event Sequence Generation

    DEFF Research Database (Denmark)

    Jensen, Casper Svenning; Prasad, Mukul R.; Møller, Anders

    2013-01-01

    of the individual event handlers of the application. The second phase builds event sequences backward from the target, using the summaries together with a UI model of the application. Our experiments on a collection of open source Android applications show that this technique can successfully produce event...

  13. Automated Testing with Targeted Event Sequence Generation

    DEFF Research Database (Denmark)

    Jensen, Casper Svenning; Prasad, Mukul R.; Møller, Anders

    2013-01-01

    of the individual event handlers of the application. The second phase builds event sequences backward from the target, using the summaries together with a UI model of the application. Our experiments on a collection of open source Android applications show that this technique can successfully produce event...

  14. Spot-Based Generations for Meta-Fibonacci Sequences

    CERN Document Server

    Dalton, Barnaby; Tanny, Stephen

    2011-01-01

    For many meta-Fibonacci sequences it is possible to identify a partition of the sequence into successive intervals (sometimes called blocks) with the property that the sequence behaves "similarly" in each block. This partition provides insights into the sequence properties. To date, for any given sequence, only ad hoc methods have been available to identify this partition. We apply a new concept - the spot-based generation sequence - to derive a general methodology for identifying this partition for a large class of meta-Fibonacci sequences. This class includes the Conolly and Conway sequences and many of their well-behaved variants, and even some highly chaotic sequences, such as Hofstadter's famous Q-sequence.

  15. The age-metallicity relation in the solar neighbourhood from a pilot sample of white dwarf-main sequence binaries

    CERN Document Server

    Rebassa-Mansergas, A; García-Berro, E; Freeman, K C; Cojocaru, R; Manser, C J; Pala, A F; Gänsicke, B T; Liu, X -W

    2016-01-01

    The age-metallicity relation (AMR) is a fundamental observational constraint for understanding how the Galactic disc formed and evolved chemically in time. However, there is not yet an agreement on the observational properties of the AMR for the solar neighbourhood, primarily due to the difficulty in obtaining accurate stellar ages for individual field stars. We have started an observational campaign for providing the much needed observational input by using wide white dwarf-main sequence (WDMS) binaries. White dwarfs are natural clocks and can be used to derive accurate ages. Metallicities can be obtained from the main sequence companions. Since the progenitors of white dwarfs and the main sequence stars were born at the same time, WDMS binaries provide a unique opportunity to observationally constrain in a robust way the properties of the AMR. In this work we present the AMR derived from analysing a pilot sample of 23 WDMS binaries and provide clear observational evidence for the lack of correlation between...

  16. On models of the genetic code generated by binary dichotomic algorithms.

    Science.gov (United States)

    Gumbel, Markus; Fimmel, Elena; Danielli, Alberto; Strüngmann, Lutz

    2015-02-01

    In this paper we introduce the concept of a BDA-generated model of the genetic code which is based on binary dichotomic algorithms (BDAs). A BDA-generated model is based on binary dichotomic algorithms (BDAs). Such a BDA partitions the set of 64 codons into two disjoint classes of size 32 each and provides a generalization of known partitions like the Rumer dichotomy. We investigate what partitions can be generated when a set of different BDAs is applied sequentially to the set of codons. The search revealed that these models are able to generate code tables with very different numbers of classes ranging from 2 to 64. We have analyzed whether there are models that map the codons to their amino acids. A perfect matching is not possible. However, we present models that describe the standard genetic code with only few errors. There are also models that map all 64 codons uniquely to 64 classes showing that BDAs can be used to identify codons precisely. This could serve as a basis for further mathematical analysis using coding theory, for example. The hypothesis that BDAs might reflect a molecular mechanism taking place in the decoding center of the ribosome is discussed. The scan demonstrated that binary dichotomic partitions are able to model different aspects of the genetic code very well. The search was performed with our tool Beady-A. This software is freely available at http://mi.informatik.hs-mannheim.de/beady-a. It requires a JVM version 6 or higher.

  17. Hunting for millimeter flares from magnetic reconnection in pre-main sequence spectroscopic binaries

    Science.gov (United States)

    Kóspál, Á.; Salter, D. M.; Hogerheijde, M. R.; Moór, A.; Blake, G. A.

    2011-03-01

    Context. Recent observations of the low-mass pre-main sequence (PMS), eccentric spectroscopic binaries DQ Tau and V773 Tau A reveal that their millimeter spectrum is occasionally dominated by flares from non-thermal emission processes. The transient activity is believed to be synchrotron in nature, resulting from powerful magnetic reconnection events when the separate magnetic structures of the binary components are briefly capable of interacting and forced to reorganize, typically near periastron. Aims: We conducted the first systematic study of the millimeter variability toward a sample of 12 PMS spectroscopic binaries with the aim to characterize the proliferation of flares amongst sources likely to experience similar interbinary reconnection events. The source sample consists entirely of short-period, close-separation binaries that possess either a high orbital eccentricity (e > 0.1) or a circular orbit (e ≈ 0). Methods: Using the MAMBO2 array on the IRAM 30 m telescope, we carried out continuous monitoring at 1.25 mm (240 GHz) over a 4-night period during which all of the high-eccentricity binaries approached periastron. We also obtained simultaneous optical VRI measurements, since a strong link is often observed between stellar reconnection events (traced via X-rays) and optical brightenings. Results: UZ Tau E is the only source to be detected at millimeter wavelengths, and it exhibited significant variation (F1.25mm = 87-179 mJy); it is also the only source to undergo strong simultaneous optical variability (ΔR ≈ 0.9 mag). The binary possesses the largest orbital eccentricity in the current sample, a predicted factor in star-star magnetic interaction events. With orbital parameters and variable accretion activity similar to DQ Tau, the millimeter behavior of UZ Tau E draws many parallels to the DQ Tau model for colliding magnetospheres. However, on the basis of our observations alone, we cannot determine whether the variability is repetitive, or if it

  18. Efficient coroutine generation of constrained Gray sequences

    CERN Document Server

    Knuth, Donald E

    2009-01-01

    We study an interesting family of cooperating coroutines, which is able to generate all patterns of bits that satisfy certain fairly general ordering constraints, changing only one bit at a time. (More precisely, the directed graph of constraints is required to be cycle-free when it is regarded as an undirected graph.) If the coroutines are implemented carefully, they yield an algorithm that needs only a bounded amount of computation per bit change, thereby solving an open problem in the field of combinatorial pattern generation.

  19. High-quality binary fringe generation via joint optimization on intensity and phase

    Science.gov (United States)

    Xiao, Yi; Li, Youfu

    2017-10-01

    There have been active studies on optimized dithering techniques to improve 3D shape measurement quality with defocused projectors. These techniques optimize the fringe quality in either phase domain or intensity domain according to their objective functions. Phase based optimization is direct and effective, but is sensitive to projector defocus levels. Intensity based optimization is robust to projector defocus levels, but it does not fully improve the phase quality. This paper presents a joint optimization technique to combine the merits of both the intensity and phase based optimization, which includes a pre-intensity optimization and a further optimization based on the synthesized error function. Then this technique is implemented in two frameworks, the whole-fringe optimization and the best-patch optimization, to generate binary fringe patterns. Both simulations and experiments show that the proposed technique can generate binary fringe patterns with high phase quality and robustness to projector defocus levels.

  20. Generation of physical map contig-specific sequences

    Directory of Open Access Journals (Sweden)

    Yanliang eJiang

    2014-07-01

    Full Text Available Rapid advances of the next-generation sequencing technologies have allowed whole genome sequencing of many species. However, with the current sequencing technologies, the whole genome sequence assemblies often fall in short in one of the four quality measurements: accuracy, contiguity, connectivity, and completeness. In particular, small-sized contigs and scaffolds limit the applicability of whole genome sequences for genetic analysis. To enhance the quality of whole genome sequence assemblies, particularly the scaffolding capabilities, additional genomic resources are required. Among these, sequences derived from known physical locations offer great powers for scaffolding. In this mini-review, we will describe the principles, procedures and applications of physical-map-derived sequences, with the focus on physical map contig-specific sequences.

  1. The first magnetic maps of a pre-main sequence binary star system - HD 155555

    CERN Document Server

    Dunstone, N J; Cameron, A Collier; Marsden, S C; Jardine, M; Stempels, H C; Vlex, J C Ramirez; Donati, J -F

    2008-01-01

    We present the first maps of the surface magnetic fields of a pre-main sequence binary system. Spectropolarimetric observations of the young, 18 Myr, HD 155555 (V824 Ara, G5IV + K0IV) system were obtained at the Anglo-Australian Telescope in 2004 and 2007. Both datasets are analysed using a new binary Zeeman Doppler imaging (ZDI) code. This allows us to simultaneously model the contribution of each component to the observed circularly polarised spectra. Stellar brightness maps are also produced for HD 155555 and compared to previous Doppler images. Our radial magnetic maps reveal a complex surface magnetic topology with mixed polarities at all latitudes. We find rings of azimuthal field on both stars, most of which are found to be non-axisymmetric with the stellar rotational axis. We also examine the field strength and the relative fraction of magnetic energy stored in the radial and azimuthal field components at both epochs. A marked weakening of the field strength of the secondary star is observed between t...

  2. RX J0942.7-7726AB: an isolated pre-main sequence wide binary

    CERN Document Server

    Murphy, Simon J; Bessell, Michael S

    2012-01-01

    We report the discovery of two young M-dwarfs, RX J0942.7-7726 (M1) and 2MASS J09424157-7727130 (M4.5), that were found only 42 arcsec apart in a survey for pre-main sequence stars surrounding the open cluster eta Chamaeleontis. Both stars have congruent proper motions and near-infrared photometry. Medium-resolution spectroscopy reveals that they are coeval (age 8-12 Myr), codistant (100-150 pc) and thus almost certainly form a true wide binary with a projected separation of 4000-6000 AU. The system appears too old and dynamically fragile to have originated in eta Cha and a traceback analysis argues for its birth in or near the Scorpius-Centaurus OB Association. RX J0942.7-7726AB joins a growing group of wide binaries kinematically linked to Sco-Cen, suggesting that such fragile systems can survive the turbulent environment of their natal molecular clouds while still being dispersed with large velocities. Conversely, the small radial velocity difference between the stars (2.7 \\pm 1.0 km/s) could mean the syst...

  3. [Automatic analysis pipeline of next-generation sequencing data].

    Science.gov (United States)

    Wenke, Li; Fengyu, Li; Siyao, Zhang; Bin, Cai; Na, Zheng; Yu, Nie; Dao, Zhou; Qian, Zhao

    2014-06-01

    The development of next-generation sequencing has generated high demand for data processing and analysis. Although there are a lot of software for analyzing next-generation sequencing data, most of them are designed for one specific function (e.g., alignment, variant calling or annotation). Therefore, it is necessary to combine them together for data analysis and to generate interpretable results for biologists. This study designed a pipeline to process Illumina sequencing data based on Perl programming language and SGE system. The pipeline takes original sequence data (fastq format) as input, calls the standard data processing software (e.g., BWA, Samtools, GATK, and Annovar), and finally outputs a list of annotated variants that researchers can further analyze. The pipeline simplifies the manual operation and improves the efficiency by automatization and parallel computation. Users can easily run the pipeline by editing the configuration file or clicking the graphical interface. Our work will facilitate the research projects using the sequencing technology.

  4. Comparison of sequence reads obtained from three next-generation sequencing platforms.

    Directory of Open Access Journals (Sweden)

    Shingo Suzuki

    Full Text Available Next-generation sequencing technologies enable the rapid cost-effective production of sequence data. To evaluate the performance of these sequencing technologies, investigation of the quality of sequence reads obtained from these methods is important. In this study, we analyzed the quality of sequence reads and SNP detection performance using three commercially available next-generation sequencers, i.e., Roche Genome Sequencer FLX System (FLX, Illumina Genome Analyzer (GA, and Applied Biosystems SOLiD system (SOLiD. A common genomic DNA sample obtained from Escherichia coli strain DH1 was applied to these sequencers. The obtained sequence reads were aligned to the complete genome sequence of E. coli DH1, to evaluate the accuracy and sequence bias of these sequence methods. We found that the fraction of "junk" data, which could not be aligned to the reference genome, was largest in the data set of SOLiD, in which about half of reads could not be aligned. Among data sets after alignment to the reference, sequence accuracy was poorest in GA data sets, suggesting relatively low fidelity of the elongation reaction in the GA method. Furthermore, by aligning the sequence reads to the E. coli strain W3110, we screened sequence differences between two E. coli strains using data sets of three different next-generation platforms. The results revealed that the detected sequence differences were similar among these three methods, while the sequence coverage required for the detection was significantly small in the FLX data set. These results provided valuable information on the quality of short sequence reads and the performance of SNP detection in three next-generation sequencing platforms.

  5. Heterogeneous Suppression of Sequential Effects in Random Sequence Generation, but Not in Operant Learning

    Science.gov (United States)

    Shteingart, Hanan; Loewenstein, Yonatan

    2016-01-01

    There is a long history of experiments in which participants are instructed to generate a long sequence of binary random numbers. The scope of this line of research has shifted over the years from identifying the basic psychological principles and/or the heuristics that lead to deviations from randomness, to one of predicting future choices. In this paper, we used generalized linear regression and the framework of Reinforcement Learning in order to address both points. In particular, we used logistic regression analysis in order to characterize the temporal sequence of participants’ choices. Surprisingly, a population analysis indicated that the contribution of the most recent trial has only a weak effect on behavior, compared to more preceding trials, a result that seems irreconcilable with standard sequential effects that decay monotonously with the delay. However, when considering each participant separately, we found that the magnitudes of the sequential effect are a monotonous decreasing function of the delay, yet these individual sequential effects are largely averaged out in a population analysis because of heterogeneity. The substantial behavioral heterogeneity in this task is further demonstrated quantitatively by considering the predictive power of the model. We show that a heterogeneous model of sequential dependencies captures the structure available in random sequence generation. Finally, we show that the results of the logistic regression analysis can be interpreted in the framework of reinforcement learning, allowing us to compare the sequential effects in the random sequence generation task to those in an operant learning task. We show that in contrast to the random sequence generation task, sequential effects in operant learning are far more homogenous across the population. These results suggest that in the random sequence generation task, different participants adopt different cognitive strategies to suppress sequential dependencies when

  6. Orbit and spin evolution of synchronous binary stars on the main sequence

    Institute of Scientific and Technical Information of China (English)

    Lin-Sen Li

    2012-01-01

    A set of synchronous equations are derived from a set of non-synchronous equations.The analytical solutions are given by solving the set of differential equations.The results of the evolutionary trend of the spin-orbit interaction are that the semi-major axis gradually shrinks with time; the orbital eccentricity gradually decreases with time until orbital circularization occurs; the orbital period gradually shortens with time and the rotational angular velocity of the primary component gradually speeds up with time before the orbit achieves circularization.The theoretical results are applied to evolution of the orbit and spin of synchronous binary stars Algol A and B that are on the main sequence.The circularization time,lifetime and the evolutionary numerical solutions of orbit and spin when circularization time occurs are estimated for Algol A and B.

  7. Generation of hierarchically correlated multivariate symbolic sequences

    CERN Document Server

    Tumminello, Mi; Mantegna, R N

    2008-01-01

    We introduce an algorithm to generate multivariate series of symbols from a finite alphabet with a given hierarchical structure of similarities. The target hierarchical structure of similarities is arbitrary, for instance the one obtained by some hierarchical clustering procedure as applied to an empirical matrix of Hamming distances. The algorithm can be interpreted as the finite alphabet equivalent of the recently introduced hierarchically nested factor model (M. Tumminello et al. EPL 78 (3) 30006 (2007)). The algorithm is based on a generating mechanism that is different from the one used in the mutation rate approach. We apply the proposed methodology for investigating the relationship between the bootstrap value associated with a node of a phylogeny and the probability of finding that node in the true phylogeny.

  8. The contribution of next generation sequencing to epilepsy genetics

    DEFF Research Database (Denmark)

    Møller, Rikke S.; Dahl, Hans A.; Helbig, Ingo

    2015-01-01

    to as epileptic encephalopathies. The increased knowledge about causative genetic variants has had a major impact on diagnosis of genetic epilepsies and has already been translated into treatment recommendations for a few genes. This article provides an overview of how next generation sequencing has advanced our......During the last decade, next generation sequencing technologies such as targeted gene panels, whole exome sequencing and whole genome sequencing have led to an explosion of gene identifications in monogenic epilepsies including both familial epilepsies and severe epilepsies, often referred...... understanding of epilepsy genetics and discusses some of the recently discovered genes in monogenic epilepsies....

  9. Short barcodes for next generation sequencing.

    Directory of Open Access Journals (Sweden)

    Katharina Mir

    Full Text Available We consider the design and evaluation of short barcodes, with a length between six and eight nucleotides, used for parallel sequencing on platforms where substitution errors dominate. Such codes should have not only good error correction properties but also the code words should fulfil certain biological constraints (experimental parameters. We compare published barcodes with codes obtained by two new constructions methods, one based on the currently best known linear codes and a simple randomized construction method. The evaluation done is with respect to the error correction capabilities, barcode size and their experimental parameters and fundamental bounds on the code size and their distance properties. We provide a list of codes for lengths between six and eight nucleotides, where for length eight, two substitution errors can be corrected. In fact, no code with larger minimum distance can exist.

  10. Short barcodes for next generation sequencing.

    Science.gov (United States)

    Mir, Katharina; Neuhaus, Klaus; Bossert, Martin; Schober, Steffen

    2013-01-01

    We consider the design and evaluation of short barcodes, with a length between six and eight nucleotides, used for parallel sequencing on platforms where substitution errors dominate. Such codes should have not only good error correction properties but also the code words should fulfil certain biological constraints (experimental parameters). We compare published barcodes with codes obtained by two new constructions methods, one based on the currently best known linear codes and a simple randomized construction method. The evaluation done is with respect to the error correction capabilities, barcode size and their experimental parameters and fundamental bounds on the code size and their distance properties. We provide a list of codes for lengths between six and eight nucleotides, where for length eight, two substitution errors can be corrected. In fact, no code with larger minimum distance can exist.

  11. Next-generation sequencing strategies for characterizing the turkey genome.

    Science.gov (United States)

    Dalloul, Rami A; Zimin, Aleksey V; Settlage, Robert E; Kim, Sungwon; Reed, Kent M

    2014-02-01

    The turkey genome sequencing project was initiated in 2008 and has relied primarily on next-generation sequencing (NGS) technologies. Our first efforts used a synergistic combination of 2 NGS platforms (Roche/454 and Illumina GAII), detailed bacterial artificial chromosome (BAC) maps, and unique assembly tools to sequence and assemble the genome of the domesticated turkey, Meleagris gallopavo. Since the first release in 2010, efforts to improve the genome assembly, gene annotation, and genomic analyses continue. The initial assembly build (2.01) represented about 89% of the genome sequence with 17X coverage depth (931 Mb). Sequence contigs were assigned to 30 of the 40 chromosomes with approximately 10% of the assembled sequence corresponding to unassigned chromosomes (ChrUn). The sequence has been refined through both genome-wide and area-focused sequencing, including shotgun and paired-end sequencing, and targeted sequencing of chromosomal regions with low or incomplete coverage. These additional efforts have improved the sequence assembly resulting in 2 subsequent genome builds of higher genome coverage (25X/Build3.0 and 30X/Build4.0) with a current sequence totaling 1,010 Mb. Further, BAC with end sequences assigned to the Z/W and MG18 (MHC) chromosomes, ChrUn, or not placed in the previous build were isolated, deeply sequenced (Hi-Seq), and incorporated into the latest build (5.0). To aid in the annotation and to generate a gene expression atlas of major tissues, a comprehensive set of RNA samples was collected at various developmental stages of female and male turkeys. Transcriptome sequencing data (using Illumina Hi-Seq) will provide information to enhance the final assembly and ultimately improve sequence annotation. The most current sequence covers more than 95% of the turkey genome and should yield a much improved gene level of annotation, making it a valuable resource for studying genetic variations underlying economically important traits in poultry.

  12. Next Generation Sequencing of Ancient DNA: Requirements, Strategies and Perspectives

    Directory of Open Access Journals (Sweden)

    Michael Knapp

    2010-07-01

    Full Text Available The invention of next-generation-sequencing has revolutionized almost all fields of genetics, but few have profited from it as much as the field of ancient DNA research. From its beginnings as an interesting but rather marginal discipline, ancient DNA research is now on its way into the centre of evolutionary biology. In less than a year from its invention next-generation-sequencing had increased the amount of DNA sequence data available from extinct organisms by several orders of magnitude. Ancient DNA  research is now not only adding a temporal aspect to evolutionary studies and allowing for the observation of evolution in real time, it also provides important data to help understand the origins of our own species. Here we review progress that has been made in next-generation-sequencing of ancient DNA over the past five years and evaluate sequencing strategies and future directions.

  13. Comparison of next generation sequencing technologies for transcriptome characterization

    Directory of Open Access Journals (Sweden)

    Soltis Douglas E

    2009-08-01

    Full Text Available Abstract Background We have developed a simulation approach to help determine the optimal mixture of sequencing methods for most complete and cost effective transcriptome sequencing. We compared simulation results for traditional capillary sequencing with "Next Generation" (NG ultra high-throughput technologies. The simulation model was parameterized using mappings of 130,000 cDNA sequence reads to the Arabidopsis genome (NCBI Accession SRA008180.19. We also generated 454-GS20 sequences and de novo assemblies for the basal eudicot California poppy (Eschscholzia californica and the magnoliid avocado (Persea americana using a variety of methods for cDNA synthesis. Results The Arabidopsis reads tagged more than 15,000 genes, including new splice variants and extended UTR regions. Of the total 134,791 reads (13.8 MB, 119,518 (88.7% mapped exactly to known exons, while 1,117 (0.8% mapped to introns, 11,524 (8.6% spanned annotated intron/exon boundaries, and 3,066 (2.3% extended beyond the end of annotated UTRs. Sequence-based inference of relative gene expression levels correlated significantly with microarray data. As expected, NG sequencing of normalized libraries tagged more genes than non-normalized libraries, although non-normalized libraries yielded more full-length cDNA sequences. The Arabidopsis data were used to simulate additional rounds of NG and traditional EST sequencing, and various combinations of each. Our simulations suggest a combination of FLX and Solexa sequencing for optimal transcriptome coverage at modest cost. We have also developed ESTcalc http://fgp.huck.psu.edu/NG_Sims/ngsim.pl, an online webtool, which allows users to explore the results of this study by specifying individualized costs and sequencing characteristics. Conclusion NG sequencing technologies are a highly flexible set of platforms that can be scaled to suit different project goals. In terms of sequence coverage alone, the NG sequencing is a dramatic advance

  14. Generating Exome Enriched Sequencing Libraries from Formalin-Fixed, Paraffin-Embedded Tissue DNA for Next-Generation Sequencing.

    Science.gov (United States)

    Marosy, Beth A; Craig, Brian D; Hetrick, Kurt N; Witmer, P Dane; Ling, Hua; Griffith, Sean M; Myers, Benjamin; Ostrander, Elaine A; Stanford, Janet L; Brody, Lawrence C; Doheny, Kimberly F

    2017-01-11

    This unit describes a technique for generating exome-enriched sequencing libraries using DNA extracted from formalin-fixed paraffin-embedded (FFPE) samples. Utilizing commercially available kits, we present a low-input FFPE workflow starting with 50 ng of DNA. This procedure includes a repair step to address damage caused by FFPE preservation that improves sequence quality. Subsequently, libraries undergo an in-solution-targeted selection for exons, followed by sequencing using the Illumina next-generation short-read sequencing platform. © 2017 by John Wiley & Sons, Inc.

  15. Next-generation sequencing and large genome assemblies

    OpenAIRE

    Henson, Joseph; Tischler, German; Ning, Zemin

    2012-01-01

    The next-generation sequencing (NGS) revolution has drastically reduced time and cost requirements for sequencing of large genomes, and also qualitatively changed the problem of assembly. This article reviews the state of the art in de novo genome assembly, paying particular attention to mammalian-sized genomes. The strengths and weaknesses of the main sequencing platforms are highlighted, leading to a discussion of assembly and the new challenges associated with NGS data. Current approaches ...

  16. Clinical Next Generation Sequencing for Precision Medicine in Cancer

    OpenAIRE

    Dong, Ling; Wang, Wanheng; Li, Alvin; Kansal, Rina; Chen, Yuhan; Hong CHEN; Li, Xinmin

    2015-01-01

    Rapid adoption of next generation sequencing (NGS) in genomic medicine has been driven by low cost, high throughput sequencing and rapid advances in our understanding of the genetic bases of human diseases. Today, the NGS method has dominated sequencing space in genomic research, and quickly entered clinical practice. Because unique features of NGS perfectly meet the clinical reality (need to do more with less), the NGS technology is becoming a driving force to realize the dream of precision ...

  17. Multiple nuclear ortholog next generation sequencing phylogeny of Daucus

    Science.gov (United States)

    Next generation sequencing is helping to solve the data insufficiency problem hindering well-resolved dominant gene phylogenies. We used Roche 454 technology to obtain DNA sequences from 93 nuclear orthologs, dispersed throughout all linkage groups of Daucus. Of these 93 orthologs, ten were designed...

  18. The SDSS spectroscopic catalogue of white dwarf-main sequence binaries: new identifications from DR9-12

    CERN Document Server

    Rebassa-Mansergas, A; Parsons, S G; Gaensicke, B T; Schreiber, M R; Garcia-Berro, E; Liu, X -W; Koester, D

    2016-01-01

    We present an updated version of the spectroscopic catalogue of white dwarf-main sequence (WDMS) binaries from the Sloan Digital Sky Survey (SDSS). We identify 939 WDMS binaries within the data releases (DR) 9-12 of SDSS plus 40 objects from DR 1-8 that we missed in our previous works, 646 of which are new. The total number of spectroscopic SDSS WDMS binaries increases to 3294. This is by far the largest and most homogeneous sample of compact binaries currently available. We use a decomposition/fitting routine to derive the stellar parameters of all systems identified here (white dwarf effective temperatures, surface gravities and masses, and secondary star spectral types). The analysis of the corresponding stellar parameter distributions shows that the SDSS WDMS binary population is seriously affected by selection effects. We also measure the NaI 8183.27, 8194.81 absorption doublet and Halpha emission radial velocities (RV) from all SDSS WDMS binary spectra identified in this work. 98 objects are found to di...

  19. Two new SB2 binaries with main sequence B-type pulsators in the Kepler field

    Science.gov (United States)

    Papics, P. I.

    2013-06-01

    OB stars are important in the chemistry and evolution of the Universe, but the sample of targets well understood from an asteroseismological point of view is still too limited to provide feedback on the current evolutionary models. Our study extends this sample with two spectroscopic binary systems. Aims. Our goal is to provide orbital solutions, fundamental parameters and abundances from disentangled high-resolution high signal-to-noise spectra, as well as to analyse and interpret the variations in the Kepler light curve of these carefully selected targets. This way we continue our efforts to map the instability strips of β Cep and slowly pulsating B stars using the combination of high-resolution ground-based spectroscopy and uninterrupted space-based photometry. Methods. We fit Keplerian orbits to radial velocities measured from selected absorption lines of high-resolution spectroscopy using synthetic composite spectra to obtain orbital solutions. We use revised masks to obtain optimal light curves from the original pixel-data from the Kepler satellite, which provided better long term stability compared to the pipeline processed light curves. We use various time-series analysis tools to explore and describe the nature of variations present in the light curve. Results. We find two eccentric double-lined spectroscopic binary systems containing a total of three main sequence B-type stars (and one F-type component) of which at least one in each system exhibits light variations. The light curve analysis (combined with spectroscopy) of the system of two B stars points towards the presence of tidally excited g modes in the primary component. We interpret the variations seen in the second system as classical g mode pulsations driven by the κ mechanism in the B type primary, and explain the unexpected power in the p mode region as a result of nonlinear resonant mode excitation.

  20. Novel Frequency Hopping Sequences Generator Based on AES Algorithm

    Institute of Scientific and Technical Information of China (English)

    李振荣; 庄奕琪; 张博; 张超

    2010-01-01

    A novel frequency hopping(FH) sequences generator based on advanced encryption standard(AES) iterated block cipher is proposed for FH communication systems.The analysis shows that the FH sequences based on AES algorithm have good performance in uniformity, correlation, complexity and security.A high-speed, low-power and low-cost ASIC of FH sequences generator is implemented by optimizing the structure of S-Box and MixColumns of AES algorithm, proposing a hierarchical power management strategy, and applying ...

  1. Quantifying population genetic differentiation from next-generation sequencing data

    DEFF Research Database (Denmark)

    Fumagalli, Matteo; Garrett Vieira, Filipe Jorge; Korneliussen, Thorfinn Sand

    2013-01-01

    Over the last few years, new high-throughput DNA sequencing technologies have dramatically increased speed and reduced sequencing costs. However, the use of these sequencing technologies is often challenged by errors and biases associated with the bioinformatical methods used for analyzing the data...... method for quantifying population genetic differentiation from next-generation sequencing data. In addition, we present a strategy to investigate population structure via Principal Components Analysis. Through extensive simulations, we compare the new method herein proposed to approaches based...... individuals, suggesting that employing this new method is useful for investigating the genetic relationships of populations sampled at low coverage....

  2. The Interior Structure Constants as an Age Diagnostic for Low-Mass, Pre-Main Sequence Detached Eclipsing Binary Stars

    CERN Document Server

    Feiden, Gregory A

    2013-01-01

    We propose a novel method for determining the ages of low-mass, pre-main sequence stellar systems using the apsidal motion of low-mass detached eclipsing binaries. The apsidal motion of a binary system with an eccentric orbit provides information regarding the interior structure constants of the individual stars. These constants are related to the normalized stellar interior density distribution and can be extracted from the predictions of stellar evolution models. We demonstrate that low-mass, pre-main sequence stars undergoing radiative core contraction display rapidly changing interior structure constants (greater than 5% per 10 Myr) that, when combined with observational determinations of the interior structure constants (with 5 -- 10% precision), allow for a robust age estimate. This age estimate, unlike those based on surface quantities, is largely insensitive to the surface layer where effects of magnetic activity are likely to be most pronounced. On the main sequence, where age sensitivity is minimal,...

  3. Minimal-Length Interoperability Test Sequences Generation via Genetic Algorithm

    Institute of Scientific and Technical Information of China (English)

    ZHONG Ning; KUANG Jing-ming; HE Zun-wen

    2008-01-01

    A novel interoperability test sequences optimization scheme is proposed in which the genetic algo-rithm(GA)is used to obtain the minimal-length interoperability test sequences.During our work,the basicin teroperability test sequences are generated based on the minimal-complete-coverage criterion,which removes the redundancy from conformance test sequences.Then interoperability sequences minimization problem can be considered as an instance of the set covering problem,and the GA is applied to remove redundancy in interoperability transitions.The results show that compared to conventional algorithm,the proposed algorithm is more practical to avoid the state space explosion problem,for it can reduce the length of the test sequences and maintain the same transition coverage.

  4. Generation of Bessel Beams at mm- and Sub mm-wavelengths by Binary Optical Elements

    Science.gov (United States)

    Yu, Y. Z.; Dou, W. B.

    2008-07-01

    In this paper, binary optical elements (BOE’s) are designed for generating Bessel beams at mm- and sub mm- wavelengths. The design tool is to combine a genetic algorithm (GA) for global optimization with a two-dimension finite-difference time-domain (2-D FDTD) method for rigorous electromagnetic computation. The design process for converting a normally incident Gaussian beam into a Bessel beam is described in detail. Numerical results demonstrate that the designed BOE’s can not only successfully produce arbitrary order Bessel beams, but also have higher diffraction efficiencies when compared with amplitude holograms.

  5. A non-hyponormal operator generating Stieltjes moment sequences

    CERN Document Server

    Jablonski, Z J; Stochel, J

    2011-01-01

    A linear operator $S$ in a complex Hilbert space $\\hh$ for which the set $\\dzn{S}$ of its $C^\\infty$-vectors is dense in $\\hh$ and $\\{\\|S^n f\\|^2\\}_{n=0}^\\infty$ is a Stieltjes moment sequence for every $f \\in \\dzn{S}$ is said to generate Stieltjes moment sequences. It is shown that there exists a closed non-hyponormal operator $S$ which generates Stieltjes moment sequences. What is more, $\\dzn{S}$ is a core of any power $S^n$ of $S$. This is established with the help of a weighted shift on a directed tree with one branching vertex. The main tool in the construction comes from the theory of indeterminate Stieltjes moment sequences. As a consequence, it is shown that there exists a non-hyponormal composition operator in an $L^2$-space (over a $\\sigma$-finite measure space) which is injective, paranormal and which generates Stieltjes moment sequences. In contrast to the case of abstract Hilbert space operators, composition operators which are formally normal and which generate Stieltjes moment sequences are alw...

  6. Third Generation Sequencing Techniques and Applications to Drug Discovery

    Science.gov (United States)

    Ozsolak, Fatih

    2012-01-01

    Introduction There is an immediate need for functional and molecular studies to decipher differences between disease and “normal” settings to identify large quantities of validated targets with the highest therapeutic utilities. Furthermore, drug mechanism of action and biomarkers to predict drug efficacy and safety need to be identified for effective design of clinical trials, decreasing attrition rates, regulatory agency approval process and drug repositioning. By expanding the power of genetics and pharmacogenetics studies, next generation nucleic acid sequencing technologies have started to play an important role in all stages of drug discovery. Areas covered This article reviews the first and second generation sequencing technologies (SGSTs) and challenges they pose to biomedicine. The article then focuses on the emerging third generation sequencing technologies (TGSTs), their technological foundations and potential contributions to drug discovery. Expert Opinion Despite the scientific and commercial success of SGSTs, the goal of rapid, comprehensive and unbiased sequencing of nucleic acids has not been achieved. TGSTs promise to increase sequencing throughput and read lengths, decrease costs, run times and error rates, eliminate biases inherent in SGSTs, and offer capabilities beyond nucleic acid sequencing. Such changes will have positive impact in all sequencing applications to drug discovery. PMID:22468954

  7. THE 2-ERROR LINEAR COMPLEXITY OF 2n-PERIODIC BINARY SEQUENCES WITH LINEAR COMPLEXITY 2n-1

    Institute of Scientific and Technical Information of China (English)

    Zhu Fengxiang; Qi Wenfeng

    2007-01-01

    Linear complexity and κ-error linear complexity of the stream cipher are two important standards to scale the randomicity of keystreams.For the 2n-periodic periodic binary sequence with linear complexity 2n-1 and κ=2,3,the number of sequences with given κ-error linear complexity and the expected κ-error linear complexity are provided.Moreover,the proportion of the sequences whose κ-error linear complexity is bigger than the expected value is analyzed.

  8. Pre-main-sequence binaries with tidally disrupted discs: the Br gamma in HD 104237

    CERN Document Server

    Garcia, P J V; Dougados, C; Bacciotti, F; Clausse, J -M; Massi, F; Mérand, A; Petrov, R; Weigelt, G

    2013-01-01

    Active pre-main-sequence binaries with separations of around ten stellar radii present a wealth of phenomena unobserved in common systems. The study of these objects is extended from Classical T Tauri stars to the Herbig Ae star HD 104237. Spectro-interferometry with the VLTI/AMBER is presented. It is found that the K-band continuum squared visibilities are compatible with a circumbinary disc with a radius of ~0.5 AU. However, a significant fraction (~50 per cent) of the flux is unresolved and not fully accounted by the stellar photospheres. The stars probably don't hold circumstellar discs, in addition to the circumbinary disk, due to the combined effects of inner magnetospheric truncation and outer tidal truncation. This unresolved flux likely arises in compact structures inside the tidally disrupted circumbinary disc. Most ($\\gtrsim 90$ per cent) of the Br gamma line emission is unresolved. The line-to-continuum spectro-astrometry shifts in time, along the direction of the Ly alpha jet known to be driven b...

  9. Estimating individual admixture proportions from next generation sequencing data

    DEFF Research Database (Denmark)

    Skotte, Line; Korneliussen, Thorfinn Sand; Albrechtsen, Anders

    2013-01-01

    Inference of population structure and individual ancestry is important both for population genetics and for association studies. With next generation sequencing technologies it is possible to obtain genetic data for all accessible genetic variations in the genome. Existing methods for admixture...... analysis rely on known genotypes. However, individual genotypes cannot be inferred from low-depth sequencing data without introducing errors. This article presents a new method for inferring an individual's ancestry that takes the uncertainty introduced in next generation sequencing data into account....... This is achieved by working directly with genotype likelihoods that contain all relevant information of the unobserved genotypes. Using simulations as well as publicly available sequencing data, we demonstrate that the presented method has great accuracy even for very low-depth data. At the same time, we...

  10. Palindromic sequence artifacts generated during next generation sequencing library preparation from historic and ancient DNA.

    Directory of Open Access Journals (Sweden)

    Bastiaan Star

    Full Text Available Degradation-specific processes and variation in laboratory protocols can bias the DNA sequence composition from samples of ancient or historic origin. Here, we identify a novel artifact in sequences from historic samples of Atlantic cod (Gadus morhua, which forms interrupted palindromes consisting of reverse complementary sequence at the 5' and 3'-ends of sequencing reads. The palindromic sequences themselves have specific properties - the bases at the 5'-end align well to the reference genome, whereas extensive misalignments exists among the bases at the terminal 3'-end. The terminal 3' bases are artificial extensions likely caused by the occurrence of hairpin loops in single stranded DNA (ssDNA, which can be ligated and amplified in particular library creation protocols. We propose that such hairpin loops allow the inclusion of erroneous nucleotides, specifically at the 3'-end of DNA strands, with the 5'-end of the same strand providing the template. We also find these palindromes in previously published ancient DNA (aDNA datasets, albeit at varying and substantially lower frequencies. This artifact can negatively affect the yield of endogenous DNA in these types of samples and introduces sequence bias.

  11. Next-Generation Sequencing: From Understanding Biology to Personalized Medicine

    Directory of Open Access Journals (Sweden)

    Benjamin Meder

    2013-03-01

    Full Text Available Within just a few years, the new methods for high-throughput next-generation sequencing have generated completely novel insights into the heritability and pathophysiology of human disease. In this review, we wish to highlight the benefits of the current state-of-the-art sequencing technologies for genetic and epigenetic research. We illustrate how these technologies help to constantly improve our understanding of genetic mechanisms in biological systems and summarize the progress made so far. This can be exemplified by the case of heritable heart muscle diseases, so-called cardiomyopathies. Here, next-generation sequencing is able to identify novel disease genes, and first clinical applications demonstrate the successful translation of this technology into personalized patient care.

  12. Exploring the switchgrass transcriptome using second-generation sequencing technology.

    Directory of Open Access Journals (Sweden)

    Yixing Wang

    Full Text Available BACKGROUND: Switchgrass (Panicum virgatum L. is a C4 perennial grass and widely popular as an important bioenergy crop. To accelerate the pace of developing high yielding switchgrass cultivars adapted to diverse environmental niches, the generation of genomic resources for this plant is necessary. The large genome size and polyploid nature of switchgrass makes whole genome sequencing a daunting task even with current technologies. Exploring the transcriptional landscape using next generation sequencing technologies provides a viable alternative to whole genome sequencing in switchgrass. PRINCIPAL FINDINGS: Switchgrass cDNA libraries from germinating seedlings, emerging tillers, flowers, and dormant seeds were sequenced using Roche 454 GS-FLX Titanium technology, generating 980,000 reads with an average read length of 367 bp. De novo assembly generated 243,600 contigs with an average length of 535 bp. Using the foxtail millet genome as a reference greatly improved the assembly and annotation of switchgrass ESTs. Comparative analysis of the 454-derived switchgrass EST reads with other sequenced monocots including Brachypodium, sorghum, rice and maize indicated a 70-80% overlap. RPKM analysis demonstrated unique transcriptional signatures of the four tissues analyzed in this study. More than 24,000 ESTs were identified in the dormant seed library. In silico analysis indicated that there are more than 2000 EST-SSRs in this collection. Expression of several orphan ESTs was confirmed by RT-PCR. SIGNIFICANCE: We estimate that about 90% of the switchgrass gene space has been covered in this analysis. This study nearly doubles the amount of EST information for switchgrass currently in the public domain. The celerity and economical nature of second-generation sequencing technologies provide an in-depth view of the gene space of complex genomes like switchgrass. Sequence analysis of closely related members of the NAD(+-malic enzyme type C4 grasses such as

  13. Implementation of Targeted Next Generation Sequencing in Clinical Diagnostics

    DEFF Research Database (Denmark)

    Larsen, Martin Jakob; Burton, Mark; Thomassen, Mads;

    Accurate mutation detection is essential in clinical genetic diagnostics of monogenic hereditary diseases. Targeted next generation sequencing (NGS) provides a promising and cost-effective alternative to Sanger sequencing and MLPA analysis currently used in most diagnostic laboratories. One...... advantage of targeted NGS is that multiple disease-specific genes can easily be sequenced simultaneously, which is favorable in genetic heterogeneous diseases. Prior to implementation in our diagnostic setting, we aimed to assess the sensitivity and specificity of targeted NGS by sequencing a collection......, respectively. For diagnostics, the sequencing coverage is essential, wherefore a minimum coverage of 30x per nucleotide in the coding regions was used as our primary quality criterion. For the majority of the included genes, we obtained adequate gene coverage, in which we were able to detect 100% of the known...

  14. Quantifying population genetic differentiation from next-generation sequencing data

    DEFF Research Database (Denmark)

    Fumagalli, Matteo; Vieira, Filipe G.; Korneliussen, Thorfinn Sand;

    2013-01-01

    Over the last few years, new high-throughput DNA sequencing technologies have dramatically increased speed and reduced sequencing costs. However, the use of these sequencing technologies is often challenged by errors and biases associated with the bioinformatical methods used for analyzing the da...... individuals, suggesting that employing this new method is useful for investigating the genetic relationships of populations sampled at low coverage....... method for quantifying population genetic differentiation from next-generation sequencing data. In addition, we present a strategy to investigate population structure via Principal Components Analysis. Through extensive simulations, we compare the new method herein proposed to approaches based...... on genotype calling and demonstrate a marked improvement in estimation accuracy for a wide range of conditions. We apply the method to a large-scale genomic data set of domesticated and wild silkworms sequenced at low coverage. We find that we can infer the fine-scale genetic structure of the sampled...

  15. Nanopore-based Fourth-generation DNA Sequencing Technology

    Institute of Scientific and Technical Information of China (English)

    Yanxiao Feng; Yuechuan Zhang; Cuifeng Ying; Deqiang Wang; Chunlei Du

    2015-01-01

    Nanopore-based sequencers, as the fourth-generation DNA sequencing technology, have the potential to quickly and reliably sequence the entire human genome for less than $1000, and possibly for even less than$100. The single-molecule techniques used by this technology allow us to further study the interaction between DNA and protein, as well as between protein and protein. Nanopore analysis opens a new door to molecular biology investigation at the single-molecule scale. In this article, we have reviewed academic achievements in nanopore technology from the past as well as the latest advances, including both biological and solid-state nanopores, and discussed their recent and potential applications.

  16. Next-Generation Sequencing and Genome Editing in Plant Virology

    OpenAIRE

    Hadidi, Ahmed; Flores, Ricardo; Candresse, Thierry; Barba, Marina

    2016-01-01

    Next-generation sequencing (NGS) has been applied to plant virology since 2009. NGS provides highly efficient, rapid, low cost DNA, or RNA high-throughput sequencing of the genomes of plant viruses and viroids and of the specific small RNAs generated during the infection process. These small RNAs, which cover frequently the whole genome of the infectious agent, are 21–24 nt long and are known as vsRNAs for viruses and vd-sRNAs for viroids. NGS has been used in a number of studies in plant vir...

  17. Next generation sequencing data of a defined microbial mock community

    Science.gov (United States)

    Singer, Esther; Andreopoulos, Bill; Bowers, Robert M.; Lee, Janey; Deshpande, Shweta; Chiniquy, Jennifer; Ciobanu, Doina; Klenk, Hans-Peter; Zane, Matthew; Daum, Christopher; Clum, Alicia; Cheng, Jan-Fang; Copeland, Alex; Woyke, Tanja

    2016-01-01

    Generating sequence data of a defined community composed of organisms with complete reference genomes is indispensable for the benchmarking of new genome sequence analysis methods, including assembly and binning tools. Moreover the validation of new sequencing library protocols and platforms to assess critical components such as sequencing errors and biases relies on such datasets. We here report the next generation metagenomic sequence data of a defined mock community (Mock Bacteria ARchaea Community; MBARC-26), composed of 23 bacterial and 3 archaeal strains with finished genomes. These strains span 10 phyla and 14 classes, a range of GC contents, genome sizes, repeat content and encompass a diverse abundance profile. Short read Illumina and long-read PacBio SMRT sequences of this mock community are described. These data represent a valuable resource for the scientific community, enabling extensive benchmarking and comparative evaluation of bioinformatics tools without the need to simulate data. As such, these data can aid in improving our current sequence data analysis toolkit and spur interest in the development of new tools. PMID:27673566

  18. Development of binary cycle generation plant (Development of 10 MW class plant)

    Energy Technology Data Exchange (ETDEWEB)

    1988-04-20

    In a binary cycle power generation system, medium/high temperature water, unutilized because of insufficient flowing force, is poured up with a DHP (Down Haul Pump) and a generator turbine is driven by an air medium obtained by heat-exchanging between the geothermal water and low boiling point medium. Merits of this system are as follows: Reduction of well drilling risk. High output obtained by a compact turbine. Enhancement of goethermal utilization. This report describes the following items. History of development (Drilling of a test well, plant design). Results in 1987 (Test well drilling, production regenerating test, reservoir analysis, plant design, natural earthwuake observation and underground water variation observation). (8 figs, 3 tabs)

  19. Enhanced method for the generation of binary Fresnel holograms based on grid-cross downsampling

    Institute of Scientific and Technical Information of China (English)

    W. K. Cheung; Peter Tsang; T. C. Poon; Changhe Zhou

    2011-01-01

    Past research has demonstrated that digital Fresnel holograms can be binarized in a non-iterative manner by downsampling the source image with a grid lattice prior to the hologram generation process. The reconstructed image of a hologram that is binarized with this approach is superior in quality compared with that obtained with direct thresholding, half-toning, and error diffusion. Despite the success, the downsampling mechanism results in a prominent texture of regularly spaced voids in the shaded regions. To alleviate this problem, an enhanced non-iterative method for the generation of binary Fresnel holograms is presented. Our method is based on a multi-direction line-sampling formed by a combined grid and cross lattice, which is capable of preserving a more solid texture in the shaded regions and enhancing the visual quality of the reconstructed image. Computer simulations and optical reconstructions are shown to demonstrate the effectiveness of our proposed technique.%Past research has demonstrated that digital Fresnel holograms can be binarized in a non-iterative manner by downsampling the source image with a grid lattice prior to the hologram generation process.The reconstructed image of a hologram that is binarized with this approach is superior in quality compared with that obtained with direct thresholding,half-toning,and error diffusion.Despite the success,the downsampling mechanism results in a prominent texture of regularly spaced voids in the shaded regions.To alleviate this problem,an enhanced non-iterative method for the generation of binary Fresnel holograms is presented.Our method is based on a multi-direction line-sampling formed by a combined grid and cross lattice,which is capable of preserving a more solid texture in the shaded regions and enhancing the visual quality of the reconstructed image.Computer simulations and optical reconstructions are shown to demonstrate the effectiveness of our proposed technique.

  20. Binary Spreading Sequences with Negative Auto-Correlation Based on Chaos Theory and Gold Sequences for Application to Asynchronous DS/CDMA Communications

    OpenAIRE

    Tsuneda, Akio; Miyazaki, Yasunori; ツネダ, アキオ; ミヤザキ, ヤスノリ; 常田, 明夫; 宮崎, 靖規

    2010-01-01

    Spreading sequences with appropriate negative auto-correlation can reduce average multiple access interference (MAI) in asynchronous DS/CDMA systems compared with the conventional Gold Sequences generated by linear feedback shift registers (LFSRs). We design spreading sequences with negative auto-correlation based on Gold sequences and the chaos theory for the Bernoulli map. By computer simulations, we evaluate BER performances of asynchronous DS/CDMA systems using the proposed sequences.

  1. Next generation sequencing (NGS): a golden tool in forensic toolkit.

    Science.gov (United States)

    Aly, S M; Sabri, D M

    2015-01-01

    The DNA analysis is a cornerstone in contemporary forensic sciences. DNA sequencing technologies are powerful tools that enrich molecular sciences in the past based on Sanger sequencing and continue to glowing these sciences based on Next generation sequencing (NGS). Next generation sequencing has excellent potential to flourish and increase the molecular applications in forensic sciences by jumping over the pitfalls of the conventional method of sequencing. The main advantages of NGS compared to conventional method that it utilizes simultaneously a large number of genetic markers with high-resolution of genetic data. These advantages will help in solving several challenges such as mixture analysis and dealing with minute degraded samples. Based on these new technologies, many markers could be examined to get important biological data such as age, geographical origins, tissue type determination, external visible traits and monozygotic twins identification. It also could get data related to microbes, insects, plants and soil which are of great medico-legal importance. Despite the dozens of forensic research involving NGS, there are requirements before using this technology routinely in forensic cases. Thus, there is a great need to more studies that address robustness of these techniques. Therefore, this work highlights the applications of forensic sciences in the era of massively parallel sequencing.

  2. A Wide-Field Survey for Transiting Hot Jupiters and Eclipsing Pre-Main-Sequence Binaries in Young Stellar Associations

    CERN Document Server

    Oelkers, Ryan J; Marshall, Jennifer L; DePoy, Darren L; Lambas, Diego G; Colazo, Carlos; Stringer, Katelyn

    2016-01-01

    The past two decades have seen a significant advancement in the detection, classification and understanding of exoplanets and binaries. This is due, in large part, to the increase in use of small-aperture telescopes (< 20 cm) to survey large areas of the sky to milli-mag precision with rapid cadence. The vast majority of the planetary and binary systems studied to date consist of main-sequence or evolved objects, leading to a dearth of knowledge of properties at early times (< 50 Myr). Only a dozen binaries and one candidate transiting Hot Jupiter are known among pre-main sequence objects, yet these are the systems that can provide the best constraints on stellar formation and planetary migration models. The deficiency in the number of well-characterized systems is driven by the inherent and aperiodic variability found in pre-main-sequence objects, which can mask and mimic eclipse signals. Hence, a dramatic increase in the number of young systems with high-quality observations is highly desirable to gui...

  3. The bioinformatics of next generation sequencing: a meeting report

    Institute of Scientific and Technical Information of China (English)

    Ravi Shankar

    2011-01-01

    @@ The Studio of Computational Biology & Bioinformatics (SCBB), IHBT, CSIR,Palampur, India organized one of the very first national workshop funded by DBT,Govt.of India, on the Bioinformatics issues associated with next generation sequencing approaches.The course structure was designed by SCBB, IHBT.The workshop took place in the IHBT premise on 17 and 18 June 2010.

  4. Next-generation sequencing approaches to understanding the oral microbiome

    NARCIS (Netherlands)

    Zaura, E.

    2012-01-01

    Until recently, the focus in dental research has been on studying a small fraction of the oral microbiome—so-called opportunistic pathogens. With the advent of next-generation sequencing (NGS) technologies, researchers now have the tools that allow for profiling of the microbiomes and metagenomes at

  5. Targeted enrichment of genomic DNA regions for next generation sequencing

    NARCIS (Netherlands)

    Mertens, F.; El-Sharawy, A.; Sauer, S.; Van Helvoort, J.; Van der Zaag, P.J.; Franke, A.; Nilsson, M.; Lehrach. H.; Brookes, A.

    2011-01-01

    In this review we discuss the latest targeted enrichment methods, and aspects of their utilization along with second generation sequencing for complex genome analysis. In doing so we provide an overview of issues involved in detecting genetic variation, for which targeted enrichment has become a pow

  6. Next-generation sequencing approaches to understanding the oral microbiome

    NARCIS (Netherlands)

    Zaura, E.

    2012-01-01

    Until recently, the focus in dental research has been on studying a small fraction of the oral microbiome—so-called opportunistic pathogens. With the advent of next-generation sequencing (NGS) technologies, researchers now have the tools that allow for profiling of the microbiomes and metagenomes at

  7. YSOVAR: SIX PRE-MAIN-SEQUENCE ECLIPSING BINARIES IN THE ORION NEBULA CLUSTER

    Energy Technology Data Exchange (ETDEWEB)

    Morales-Calderon, M.; Stauffer, J. R.; Rebull, L. M. [Spitzer Science Center, California Institute of Technology, 1200 E California Blvd., Pasadena, CA 91125 (United States); Stassun, K. G. [Physics and Astronomy Department, Vanderbilt University, 1807 Station B, Nashville, TN 37235 (United States); Vrba, F. J. [U. S. Naval Observatory, Flagstaff Station, 10391 W. Naval Observatory Road, Flagstaff, AZ 86001-8521 (United States); Prato, L. [Lowell Observatory, 1400 West Mars Hill Road, Flagstaff, AZ 86001 (United States); Hillenbrand, L. A.; Carpenter, J. M. [Astronomy Department, California Institute of Technology, 1200 E California Blvd., Pasadena, CA 91125 (United States); Terebey, S.; Angione, J. [Department of Physics and Astronomy, California State University at Los Angeles, Los Angeles, CA 90032 (United States); Covey, K. R. [Department of Astronomy, Cornell University, 226 Space Sciences Building, Ithaca, NY 14853 (United States); Terndrup, D. M. [Department of Astronomy, The Ohio State University, 140 West 18th Avenue, Columbus, OH 43210 (United States); Gutermuth, R. [Department of Astronomy, University of Massachusetts, Amherst, MA 01003 (United States); Song, I. [Physics and Astronomy Department, University of Georgia, Athens, GA 30602-2451 (United States); Plavchan, P. [NASA Exoplanet Science Institute, California Institute of Technology, Pasadena, CA 91125 (United States); Marchis, F. [SETI Institute, Carl Sagan Center, 189 N San Bernado Av, Mountain View, CA 94043 (United States); Garcia, E. V. [Department of Physics, Fisk University, 1000 17th Ave. N, Nashville, TN 37208 (United States); Margheim, S. [Gemini Observatory, Southern Operations Center, Casilla 603, La Serena (Chile); Luhman, K. L. [Department of Astronomy and Astrophysics, The Pennsylvania State University, University Park, PA 16802 (United States); Irwin, J. M., E-mail: mariamc@cab.inta-csic.es [Harvard-Smithsonian Center for Astrophysics, 60 Garden St., Cambridge, MA 02138 (United States)

    2012-07-10

    Eclipsing binaries (EBs) provide critical laboratories for empirically testing predictions of theoretical models of stellar structure and evolution. Pre-main-sequence (PMS) EBs are particularly valuable, both due to their rarity and the highly dynamic nature of PMS evolution, such that a dense grid of PMS EBs is required to properly calibrate theoretical PMS models. Analyzing multi-epoch, multi-color light curves for {approx}2400 candidate Orion Nebula Cluster (ONC) members from our Warm Spitzer Exploration Science Program YSOVAR, we have identified 12 stars whose light curves show eclipse features. Four of these 12 EBs are previously known. Supplementing our light curves with follow-up optical and near-infrared spectroscopy, we establish two of the candidates as likely field EBs lying behind the ONC. We confirm the remaining six candidate systems, however, as newly identified ONC PMS EBs. These systems increase the number of known PMS EBs by over 50% and include the highest mass ({theta}{sup 1} Ori E, for which we provide a complete set of well-determined parameters including component masses of 2.807 and 2.797 M{sub Sun }) and longest-period (ISOY J053505.71-052354.1, P {approx} 20 days) PMS EBs currently known. In two cases ({theta}{sup 1} Ori E and ISOY J053526.88-044730.7), enough photometric and spectroscopic data exist to attempt an orbit solution and derive the system parameters. For the remaining systems, we combine our data with literature information to provide a preliminary characterization sufficient to guide follow-up investigations of these rare, benchmark systems.

  8. Next-generation sequencing and large genome assemblies.

    Science.gov (United States)

    Henson, Joseph; Tischler, German; Ning, Zemin

    2012-06-01

    The next-generation sequencing (NGS) revolution has drastically reduced time and cost requirements for sequencing of large genomes, and also qualitatively changed the problem of assembly. This article reviews the state of the art in de novo genome assembly, paying particular attention to mammalian-sized genomes. The strengths and weaknesses of the main sequencing platforms are highlighted, leading to a discussion of assembly and the new challenges associated with NGS data. Current approaches to assembly are outlined and the various software packages available are introduced and compared. The question of whether quality assemblies can be produced using short-read NGS data alone, or whether it must be combined with more expensive sequencing techniques, is considered. Prospects for future assemblers and tests of assembly performance are also discussed.

  9. Using chaos to generate variations on movement sequences

    Science.gov (United States)

    Bradley, Elizabeth; Stuart, Joshua

    1998-12-01

    We describe a method for introducing variations into predefined motion sequences using a chaotic symbol-sequence reordering technique. A progression of symbols representing the body positions in a dance piece, martial arts form, or other motion sequence is mapped onto a chaotic trajectory, establishing a symbolic dynamics that links the movement sequence and the attractor structure. A variation on the original piece is created by generating a trajectory with slightly different initial conditions, inverting the mapping, and using special corpus-based graph-theoretic interpolation schemes to smooth any abrupt transitions. Sensitive dependence guarantees that the variation is different from the original; the attractor structure and the symbolic dynamics guarantee that the two resemble one another in both aesthetic and mathematical senses.

  10. Application of next generation sequencing technology in Mendelian movement disorders.

    Science.gov (United States)

    Wang, Yumin; Pan, Xuya; Xue, Dan; Li, Yuwei; Zhang, Xueying; Kuang, Biao; Zheng, Jiabo; Deng, Hao; Li, Xiaoling; Xiong, Wei; Zeng, Zhaoyang; Li, Guiyuan

    2016-02-01

    Next generation sequencing (NGS) has developed very rapidly in the last decade. Compared with Sanger sequencing, NGS has the advantages of high sensitivity and high throughput. Movement disorders are a common type of neurological disease. Although traditional linkage analysis has become a standard method to identify the pathogenic genes in diseases, it is getting difficult to find new pathogenic genes in rare Mendelian disorders, such as movement disorders, due to a lack of appropriate families with high penetrance or enough affected individuals. Thus, NGS is an ideal approach to identify the causal alleles for inherited disorders. NGS is used to identify genes in several diseases and new mutant sites in Mendelian movement disorders. This article reviewed the recent progress in NGS and the use of NGS in Mendelian movement disorders from genome sequencing and transcriptome sequencing. A perspective on how NGS could be employed in rare Mendelian disorders is also provided.

  11. Neutrino-driven explosions of ultra-stripped type Ic supernovae generating binary neutron stars

    CERN Document Server

    Suwa, Yudai; Shibata, Masaru; Umeda, Hideyuki; Takahashi, Koh

    2015-01-01

    We study explosion characteristics of ultra-stripped supernovae (SNe), which are candidates of SNe generating binary neutron stars (NSs). As a first step, we perform stellar evolutionary simulations of bare carbon-oxygen cores of mass from 1.45 to 2.0 $M_\\odot$ until the iron cores become unstable and start collapsing. We then perform axisymmetric hydrodynamics simulations with spectral neutrino transport using these stellar evolution outcomes as initial conditions. All models exhibit successful explosions driven by neutrino heating. The diagnostic explosion energy, ejecta mass, Ni mass, and NS mass are typically $\\sim 10^{50}$ erg, $\\sim 0.1 M_\\odot$, $\\sim 0.01M_\\odot$, and $\\approx 1.3 M_\\odot$, which are compatible with observations of rapidly-evolving and luminous transient such as SN 2005ek. We also find that the ultra-stripped SN is a candidate for producing the secondary low-mass NS in the observed compact binary NSs like PSR J0737-3039.

  12. Next generation sequencing reveals the hidden diversity of zooplankton assemblages.

    Directory of Open Access Journals (Sweden)

    Penelope K Lindeque

    Full Text Available BACKGROUND: Zooplankton play an important role in our oceans, in biogeochemical cycling and providing a food source for commercially important fish larvae. However, difficulties in correctly identifying zooplankton hinder our understanding of their roles in marine ecosystem functioning, and can prevent detection of long term changes in their community structure. The advent of massively parallel next generation sequencing technology allows DNA sequence data to be recovered directly from whole community samples. Here we assess the ability of such sequencing to quantify richness and diversity of a mixed zooplankton assemblage from a productive time series site in the Western English Channel. METHODOLOGY/PRINCIPLE FINDINGS: Plankton net hauls (200 µm were taken at the Western Channel Observatory station L4 in September 2010 and January 2011. These samples were analysed by microscopy and metagenetic analysis of the 18S nuclear small subunit ribosomal RNA gene using the 454 pyrosequencing platform. Following quality control a total of 419,041 sequences were obtained for all samples. The sequences clustered into 205 operational taxonomic units using a 97% similarity cut-off. Allocation of taxonomy by comparison with the National Centre for Biotechnology Information database identified 135 OTUs to species level, 11 to genus level and 1 to order, <2.5% of sequences were classified as unknowns. By comparison a skilled microscopic analyst was able to routinely enumerate only 58 taxonomic groups. CONCLUSIONS: Metagenetics reveals a previously hidden taxonomic richness, especially for Copepoda and hard-to-identify meroplankton such as Bivalvia, Gastropoda and Polychaeta. It also reveals rare species and parasites. We conclude that Next Generation Sequencing of 18S amplicons is a powerful tool for elucidating the true diversity and species richness of zooplankton communities. While this approach allows for broad diversity assessments of plankton it may

  13. Next Generation Sequencing of Pooled Samples: Guideline for Variants’ Filtering

    Science.gov (United States)

    Anand, Santosh; Mangano, Eleonora; Barizzone, Nadia; Bordoni, Roberta; Sorosina, Melissa; Clarelli, Ferdinando; Corrado, Lucia; Martinelli Boneschi, Filippo; D’Alfonso, Sandra; De Bellis, Gianluca

    2016-01-01

    Sequencing large number of individuals, which is often needed for population genetics studies, is still economically challenging despite falling costs of Next Generation Sequencing (NGS). Pool-seq is an alternative cost- and time-effective option in which DNA from several individuals is pooled for sequencing. However, pooling of DNA creates new problems and challenges for accurate variant call and allele frequency (AF) estimation. In particular, sequencing errors confound with the alleles present at low frequency in the pools possibly giving rise to false positive variants. We sequenced 996 individuals in 83 pools (12 individuals/pool) in a targeted re-sequencing experiment. We show that Pool-seq AFs are robust and reliable by comparing them with public variant databases and in-house SNP-genotyping data of individual subjects of pools. Furthermore, we propose a simple filtering guideline for the removal of spurious variants based on the Kolmogorov-Smirnov statistical test. We experimentally validated our filters by comparing Pool-seq to individual sequencing data showing that the filters remove most of the false variants while retaining majority of true variants. The proposed guideline is fairly generic in nature and could be easily applied in other Pool-seq experiments. PMID:27670852

  14. Modelling Nonlinear Sequence Generators in terms of Linear Cellular Automata

    CERN Document Server

    Fúster-Sabater, Amparo; 10.1016/j.apm.2005.08.013

    2010-01-01

    In this work, a wide family of LFSR-based sequence generators, the so-called Clock-Controlled Shrinking Generators (CCSGs), has been analyzed and identified with a subset of linear Cellular Automata (CA). In fact, a pair of linear models describing the behavior of the CCSGs can be derived. The algorithm that converts a given CCSG into a CA-based linear model is very simple and can be applied to CCSGs in a range of practical interest. The linearity of these cellular models can be advantageously used in two different ways: (a) for the analysis and/or cryptanalysis of the CCSGs and (b) for the reconstruction of the output sequence obtained from this kind of generators.

  15. Generation of Physical Map Contig-Specific Sequences Useful for Whole Genome Sequence Scaffolding

    Science.gov (United States)

    Jiang, Yanliang; Ninwichian, Parichart; Liu, Shikai; Zhang, Jiaren; Kucuktas, Huseyin; Sun, Fanyue; Kaltenboeck, Ludmilla; Sun, Luyang; Bao, Lisui; Liu, Zhanjiang

    2013-01-01

    Along with the rapid advances of the nextgen sequencing technologies, more and more species are added to the list of organisms whose whole genomes are sequenced. However, the assembled draft genome of many organisms consists of numerous small contigs, due to the short length of the reads generated by nextgen sequencing platforms. In order to improve the assembly and bring the genome contigs together, more genome resources are needed. In this study, we developed a strategy to generate a valuable genome resource, physical map contig-specific sequences, which are randomly distributed genome sequences in each physical contig. Two-dimensional tagging method was used to create specific tags for 1,824 physical contigs, in which the cost was dramatically reduced. A total of 94,111,841 100-bp reads and 315,277 assembled contigs are identified containing physical map contig-specific tags. The physical map contig-specific sequences along with the currently available BAC end sequences were then used to anchor the catfish draft genome contigs. A total of 156,457 genome contigs (~79% of whole genome sequencing assembly) were anchored and grouped into 1,824 pools, in which 16,680 unique genes were annotated. The physical map contig-specific sequences are valuable resources to link physical map, genetic linkage map and draft whole genome sequences, consequently have the capability to improve the whole genome sequences assembly and scaffolding, and improve the genome-wide comparative analysis as well. The strategy developed in this study could also be adopted in other species whose whole genome assembly is still facing a challenge. PMID:24205335

  16. Generation of physical map contig-specific sequences useful for whole genome sequence scaffolding.

    Directory of Open Access Journals (Sweden)

    Yanliang Jiang

    Full Text Available Along with the rapid advances of the nextgen sequencing technologies, more and more species are added to the list of organisms whose whole genomes are sequenced. However, the assembled draft genome of many organisms consists of numerous small contigs, due to the short length of the reads generated by nextgen sequencing platforms. In order to improve the assembly and bring the genome contigs together, more genome resources are needed. In this study, we developed a strategy to generate a valuable genome resource, physical map contig-specific sequences, which are randomly distributed genome sequences in each physical contig. Two-dimensional tagging method was used to create specific tags for 1,824 physical contigs, in which the cost was dramatically reduced. A total of 94,111,841 100-bp reads and 315,277 assembled contigs are identified containing physical map contig-specific tags. The physical map contig-specific sequences along with the currently available BAC end sequences were then used to anchor the catfish draft genome contigs. A total of 156,457 genome contigs (~79% of whole genome sequencing assembly were anchored and grouped into 1,824 pools, in which 16,680 unique genes were annotated. The physical map contig-specific sequences are valuable resources to link physical map, genetic linkage map and draft whole genome sequences, consequently have the capability to improve the whole genome sequences assembly and scaffolding, and improve the genome-wide comparative analysis as well. The strategy developed in this study could also be adopted in other species whose whole genome assembly is still facing a challenge.

  17. Temporally consistent virtual camera generation from stereo image sequences

    Science.gov (United States)

    Fox, Simon R.; Flack, Julien; Shao, Juliang; Harman, Phil

    2004-05-01

    The recent emergence of auto-stereoscopic 3D viewing technologies has increased demand for the creation of 3D video content. A range of glasses-free multi-viewer screens have been developed that require as many as 9 views generated for each frame of video. This presents difficulties in both view generation and transmission bandwidth. This paper examines the use of stereo video capture as a means to generate multiple scene views via disparity analysis. A machine learning approach is applied to learn relationships between disparity generated depth information and source footage, and to generate depth information in a temporally smooth manner for both left and right eye image sequences. A view morphing approach to multiple view rendering is described which provides an excellent 3D effect on a range of glasses-free displays, while providing robustness to inaccurate stereo disparity calculations.

  18. Next generation sequencing in sporadic retinoblastoma patients reveals somatic mosaicism.

    Science.gov (United States)

    Amitrano, Sara; Marozza, Annabella; Somma, Serena; Imperatore, Valentina; Hadjistilianou, Theodora; De Francesco, Sonia; Toti, Paolo; Galimberti, Daniela; Meloni, Ilaria; Cetta, Francesco; Piu, Pietro; Di Marco, Chiara; Dosa, Laura; Lo Rizzo, Caterina; Carignani, Giulia; Mencarelli, Maria Antonietta; Mari, Francesca; Renieri, Alessandra; Ariani, Francesca

    2015-11-01

    In about 50% of sporadic cases of retinoblastoma, no constitutive RB1 mutations are detected by conventional methods. However, recent research suggests that, at least in some of these cases, there is somatic mosaicism with respect to RB1 normal and mutant alleles. The increased availability of next generation sequencing improves our ability to detect the exact percentage of patients with mosaicism. Using this technology, we re-tested a series of 40 patients with sporadic retinoblastoma: 10 of them had been previously classified as constitutional heterozygotes, whereas in 30 no RB1 mutations had been found in lymphocytes. In 3 of these 30 patients, we have now identified low-level mosaic variants, varying in frequency between 8 and 24%. In 7 out of the 10 cases previously classified as heterozygous from testing blood cells, we were able to test additional tissues (ocular tissues, urine and/or oral mucosa): in three of them, next generation sequencing has revealed mosaicism. Present results thus confirm that a significant fraction (6/40; 15%) of sporadic retinoblastoma cases are due to postzygotic events and that deep sequencing is an efficient method to unambiguously distinguish mosaics. Re-testing of retinoblastoma patients through next generation sequencing can thus provide new information that may have important implications with respect to genetic counseling and family care.

  19. Impact of Next Generation Sequencing Techniques in Food Microbiology

    Science.gov (United States)

    Mayo, Baltasar; Rachid, Caio T. C. C; Alegría, Ángel; Leite, Analy M. O; Peixoto, Raquel S; Delgado, Susana

    2014-01-01

    Understanding the Maxam-Gilbert and Sanger sequencing as the first generation, in recent years there has been an explosion of newly-developed sequencing strategies, which are usually referred to as next generation sequencing (NGS) techniques. NGS techniques have high-throughputs and produce thousands or even millions of sequences at the same time. These sequences allow for the accurate identification of microbial taxa, including uncultivable organisms and those present in small numbers. In specific applications, NGS provides a complete inventory of all microbial operons and genes present or being expressed under different study conditions. NGS techniques are revolutionizing the field of microbial ecology and have recently been used to examine several food ecosystems. After a short introduction to the most common NGS systems and platforms, this review addresses how NGS techniques have been employed in the study of food microbiota and food fermentations, and discusses their limits and perspectives. The most important findings are reviewed, including those made in the study of the microbiota of milk, fermented dairy products, and plant-, meat- and fish-derived fermented foods. The knowledge that can be gained on microbial diversity, population structure and population dynamics via the use of these technologies could be vital in improving the monitoring and manipulation of foods and fermented food products. They should also improve their safety. PMID:25132799

  20. Sequencing of BAC pools by different next generation sequencing platforms and strategies

    Directory of Open Access Journals (Sweden)

    Scholz Uwe

    2011-10-01

    Full Text Available Abstract Background Next generation sequencing of BACs is a viable option for deciphering the sequence of even large and highly repetitive genomes. In order to optimize this strategy, we examined the influence of read length on the quality of Roche/454 sequence assemblies, to what extent Illumina/Solexa mate pairs (MPs improve the assemblies by scaffolding and whether barcoding of BACs is dispensable. Results Sequencing four BACs with both FLX and Titanium technologies revealed similar sequencing accuracy, but showed that the longer Titanium reads produce considerably less misassemblies and gaps. The 454 assemblies of 96 barcoded BACs were improved by scaffolding 79% of the total contig length with MPs from a non-barcoded library. Assembly of the unmasked 454 sequences without separation by barcodes revealed chimeric contig formation to be a major problem, encompassing 47% of the total contig length. Masking the sequences reduced this fraction to 24%. Conclusion Optimal BAC pool sequencing should be based on the longest available reads, with barcoding essential for a comprehensive assessment of both repetitive and non-repetitive sequence information. When interest is restricted to non-repetitive regions and repeats are masked prior to assembly, barcoding is non-essential. In any case, the assemblies can be improved considerably by scaffolding with non-barcoded BAC pool MPs.

  1. GENERATION OF COMPLEMENTARY CODES AND DOUBLY CO-OPERATIVE TERNARY SEQUENCES AND THEIR COMPARITIVE STUDY IN AMBIGUITY DOMAIN

    Directory of Open Access Journals (Sweden)

    K. SRIHARI RAO,

    2011-03-01

    Full Text Available Pulse coding and linear frequency modulation in analog or discrete form are widely used in radar systems for pulse compression to achieve high range resolution. Such coded waveforms are detected by a threshold detector after matched filtering. Complementary codes, which use multiple binary sequences exhibit the most desirable characteristics, so that they achieve perfect side lobe cancellation.Here generation of complementary codes is presented whose sum of autocorrelation functions is double the length of the sequence for zero shift and zero for other shifts and pairs of doubly co-operative ternary sequences which are co-operative not only in the utocorrelation domain but also in the signal domain is presented. Theenergy efficiency of the sequences is found as unity or 100%. The hoice of the selection of the sequence in the case of complementary sequences is restricted to a few number of sequences where as in doubly co-operative sequences, the choice is broader. This is due to the property of the co-operative sequence is slightly relaxedfrom complementary codes as side lobe is relaxed to ±1 from zero in the case of complementary codes. The radar ambiguity function epresents the output of the matched filter used by the radar esigners which provides information about how different waveforms may be suitable for various radar applications. The behaviour of complementary sequences and doubly co-operative sequences is studied in ambiguity domain.

  2. Two new SB2 binaries with main sequence B-type pulsators in the Kepler field

    Science.gov (United States)

    Pápics, P. I.; Tkachenko, A.; Aerts, C.; Briquet, M.; Marcos-Arenal, P.; Beck, P. G.; Uytterhoeven, K.; Triviño Hage, A.; Southworth, J.; Clubb, K. I.; Bloemen, S.; Degroote, P.; Jackiewicz, J.; McKeever, J.; Van Winckel, H.; Niemczura, E.; Gameiro, J. F.; Debosscher, J.

    2013-05-01

    Context. OB stars are important in the chemistry and evolution of the Universe, but the sample of targets that is well understood from an asteroseismological point of view is still too limited to provide feedback on the current evolutionary models. Aims: We extend this sample with two spectroscopic binary systems. Our goal is to provide orbital solutions, fundamental parameters, and abundances from disentangled high-resolution high signal-to-noise spectra, as well as to analyse and interpret the variations in the Kepler light curve of these carefully selected targets. This way we continue our efforts to map the instability strips of β Cep and slowly pulsating B stars using the combination of high-resolution ground-based spectroscopy and uninterrupted space-based photometry. Methods: We fit Keplerian orbits to radial velocities measured from selected absorption lines of high-resolution spectroscopy using synthetic composite spectra to obtain orbital solutions. We used revised masks to obtain optimal light curves from the original pixel-data from the Kepler satellite, which provided better long-term stability compared to the pipeline-processed light curves. We used various time-series analysis tools to explore and describe the nature of variations present in the light curve. Results: We find two eccentric double-lined spectroscopic binary systems containing a total of three main sequence B-type stars (and one F-type component), of which at least one in each system exhibits light variations. The light curve analysis (combined with spectroscopy) of the system of two B stars points towards the presence of tidally excited g modes in the primary component. We interpret the variations seen in the second system as classical g mode pulsations driven by the κ mechanism in the B type primary, and explain the unexpected power in the p mode region as the result of nonlinear resonant mode excitation. Based on observations made with the Mercator telescope, operated by the

  3. An M Dwarf Companion to an F-type Star in a Young Main-sequence Binary

    Science.gov (United States)

    Eigmüller, Ph.; Eislöffel, J.; Csizmadia, Sz.; Lehmann, H.; Erikson, A.; Fridlund, M.; Hartmann, M.; Hatzes, A.; Pasternacki, Th.; Rauer, H.; Tkachenko, A.; Voss, H.

    2016-03-01

    Only a few well characterized very low-mass M dwarfs are known today. Our understanding of M dwarfs is vital as these are the most common stars in our solar neighborhood. We aim to characterize the properties of a rare F+dM stellar system for a better understanding of the low-mass end of the Hertzsprung-Russel diagram. We used photometric light curves and radial velocity follow-up measurements to study the binary. Spectroscopic analysis was used in combination with isochrone fitting to characterize the primary star. The primary star is an early F-type main-sequence star with a mass of (1.493 ± 0.073) M⊙ and a radius of (1.474 ± 0.040) R⊙. The companion is an M dwarf with a mass of (0.188 ± 0.014) M⊙ and a radius of (0.234 ± 0.009) R⊙. The orbital period is (1.35121 ± 0.00001) days. The secondary star is among the lowest-mass M dwarfs known to date. The binary has not reached a 1:1 spin-orbit synchronization. This indicates a young main-sequence binary with an age below ˜250 Myr. The mass-radius relation of both components are in agreement with this finding.

  4. AN M DWARF COMPANION TO AN F-TYPE STAR IN A YOUNG MAIN-SEQUENCE BINARY

    Energy Technology Data Exchange (ETDEWEB)

    Eigmüller, Ph.; Csizmadia, Sz.; Erikson, A.; Fridlund, M.; Pasternacki, Th.; Rauer, H. [Institute of Planetary Research, German Aerospace Center Rutherfordstr. 2, D-12489 Berlin (Germany); Eislöffel, J.; Lehmann, H.; Hartmann, M.; Hatzes, A. [Thüringer Landessternwarte Tautenburg Sternwarte 5, D-07778 Tautenburg (Germany); Tkachenko, A. [Instituut voor Sterrenkunde, KU Leuven Celestijnenlaan 200D, 3001 Leuven (Belgium); Voss, H., E-mail: philipp.eigmueller@dlr.de [Universitat de Barcelona, Department of Astronomy and Meteorology Martí i Franquès, 1, E-08028 Barcelona (Spain)

    2016-03-15

    Only a few well characterized very low-mass M dwarfs are known today. Our understanding of M dwarfs is vital as these are the most common stars in our solar neighborhood. We aim to characterize the properties of a rare F+dM stellar system for a better understanding of the low-mass end of the Hertzsprung–Russel diagram. We used photometric light curves and radial velocity follow-up measurements to study the binary. Spectroscopic analysis was used in combination with isochrone fitting to characterize the primary star. The primary star is an early F-type main-sequence star with a mass of (1.493 ± 0.073) M{sub ⊙} and a radius of (1.474 ± 0.040) R{sub ⊙}. The companion is an M dwarf with a mass of (0.188 ± 0.014) M{sub ⊙} and a radius of (0.234 ± 0.009) R{sub ⊙}. The orbital period is (1.35121 ± 0.00001) days. The secondary star is among the lowest-mass M dwarfs known to date. The binary has not reached a 1:1 spin–orbit synchronization. This indicates a young main-sequence binary with an age below ∼250 Myr. The mass–radius relation of both components are in agreement with this finding.

  5. An M dwarf Companion to an F-type Star in a young main-sequence binary

    CERN Document Server

    Eigmüller, Ph; Csizmadia, Sz; Lehmann, H; Erikson, A; Fridlund, M; Hartmann, M; Hatzes, A; Pasternacki, Th; Rauer, H; Tkachenko, A; Voss, H

    2016-01-01

    Only a few well characterized very low-mass M dwarfs are known today. Our understanding of M dwarfs is vital as these are the most common stars in our solar neighborhood. We aim to characterize the properties of a rare F+dM stellar system for a better understanding of the low-mass end of the Hertzsprung-Russel diagram. We used photometric light curves and radial velocity follow-up measurements to study the binary. Spectro- scopic analysis was used in combination with isochrone fitting to characterize the primary star. The primary star is an early F-type main-sequence star with a mass of (1.493 +- 0.073) Msun and a radius of (1.474 +- 0.040) Rsun. The companion is an M dwarf with a mass of (0.188 +- 0.014) Msun and a radius of (0.234 +- 0.009) Rsun. The orbital period is (1.35121 +- 0:00001)d. The secondary star is among the lowest-mass M dwarfs known to date. The binary has not reached a 1:1 spin-orbit synchronization. This indicates a young main-sequence binary with an age below ~250 Myrs. The mass-radius re...

  6. KNIME4NGS: a comprehensive toolbox for Next Generation Sequencing analysis.

    Science.gov (United States)

    Hastreiter, Maximilian; Jeske, Tim; Hoser, Jonathan; Kluge, Michael; Ahomaa, Kaarin; Friedl, Marie-Sophie; Kopetzky, Sebastian J; Quell, Jan-Dominik; Werner Mewes, H-; Küffner, Robert

    2017-01-09

    Analysis of Next Generation Sequencing (NGS) data requires the processing of large datasets by chaining various tools with complex input and output formats. In order to automate data analysis, we propose to standardize NGS tasks into modular workflows. This simplifies reliable handling and processing of NGS data, and corresponding solutions become substantially more reproducible and easier to maintain. Here, we present a documented, linux-based, toolbox of 42 processing modules that are combined to construct workflows facilitating a variety of tasks such as DNAseq and RNAseq analysis. We also describe important technical extensions. The high throughput executor (HTE) helps to increase the reliability and to reduce manual interventions when processing complex datasets. We also provide a dedicated binary manager that assists users in obtaining the modules' executables and keeping them up to date. As basis for this actively developed toolbox we use the workflow management software KNIME.

  7. Understanding Cancer Genome and Its Evolution by Next Generation Sequencing

    DEFF Research Database (Denmark)

    Hou, Yong

    Cancer will cause 13 million deaths by the year of 2030, ranking the second leading cause of death worldwide. Previous studies indicate that most of the cancers originate from cells that acquired somatic mutations and evolved as Darwin Theory. Ten biological insights of cancer have been summarized...... recently. Cutting-age technologies like next generation sequencing (NGS) enable exploring cancer genome and evolution much more efficiently. However, integrated cancer genome sequencing studies showed great inter-/intra-tumoral heterogeneity (ITH) and complex evolution patterns beyond the cancer biological...... evolution by NGS, we first developed high throughput single cell sequencing (SCS) pipeline on whole exome and trascriptome and updated the pipeline after systematically reviewed the existed single cell whole genome amplification (WGA) and whole transcriptome amplification methods. Using SCS pipeline we...

  8. Application of Next Generation Sequencing on Genetic Testing

    DEFF Research Database (Denmark)

    Li, Jian

    The discovery of genetic factors behind increasing number of human diseases and the growth of education of genetic knowledge to the public make demands for genetic testing increase rapidly. However, traditional genetic testing methods cannot meet all kinds of the requirements. Next generation...... sequencing (NGS) featured with high throughput and low cost of sequencing capacity develops fast, especially with the improvement of its read length, read accuracy and the immergence of small-sized machines, making it a powerful genetic testing tool. In this study, we applied NGS to develop novel genetic...... developed a targeted sequencing based preimplantation genetic diagnosis (PGD) method for monogenic diseases and tested it in a family suffering from β-thalassaemia major undergoing PGD. Moreover, we developed a method which can achieve detection of point mutation and copy number variation simultaneously...

  9. Application of Next Generation Sequencing on Genetic Testing

    DEFF Research Database (Denmark)

    Li, Jian

    The discovery of genetic factors behind increasing number of human diseases and the growth of education of genetic knowledge to the public make demands for genetic testing increase rapidly. However, traditional genetic testing methods cannot meet all kinds of the requirements. Next generation...... sequencing (NGS) featured with high throughput and low cost of sequencing capacity develops fast, especially with the improvement of its read length, read accuracy and the immergence of small-sized machines, making it a powerful genetic testing tool. In this study, we applied NGS to develop novel genetic...... developed a targeted sequencing based preimplantation genetic diagnosis (PGD) method for monogenic diseases and tested it in a family suffering from β-thalassaemia major undergoing PGD. Moreover, we developed a method which can achieve detection of point mutation and copy number variation simultaneously...

  10. MPD: multiplex primer design for next-generation targeted sequencing.

    Science.gov (United States)

    Wingo, Thomas S; Kotlar, Alex; Cutler, David J

    2017-01-05

    Targeted resequencing offers a cost-effective alternative to whole-genome and whole-exome sequencing when investigating regions known to be associated with a trait or disease. There are a number of approaches to targeted resequencing, including microfluidic PCR amplification, which may be enhanced by multiplex PCR. Currently, there is no open-source software that can design next-generation multiplex PCR experiments that ensures primers are unique at a genome-level and efficiently pools compatible primers. We present MPD, a software package that automates the design of multiplex PCR primers for next-generation sequencing. The core of MPD is implemented in C for speed and uses a hashed genome to ensure primer uniqueness, avoids placing primers over sites of known variation, and efficiently pools compatible primers. A JavaScript web application ( http://multiplexprimer.io ) utilizing the MPD Perl package provides a convenient platform for users to make designs. Using a realistic set of genes identified by genome-wide association studies (GWAS), we achieve 90% coverage of all exonic regions using stringent design criteria. Using the first 47 primer pools for wet-lab validation, we sequenced ~25Kb at 99.7% completeness with a mean coverage of 300X among 313 samples simultaneously and identified 224 variants. The number and nature of variants we observe are consistent with high quality sequencing. MPD can successfully design multiplex PCR experiments suitable for next-generation sequencing, and simplifies retooling targeted resequencing pipelines to focus on new targets as new genetic evidence emerges.

  11. All-optical pseudorandom bit sequences generator based on TOADs

    Science.gov (United States)

    Sun, Zhenchao; Wang, Zhi; Wu, Chongqing; Wang, Fu; Li, Qiang

    2016-03-01

    A scheme for all-optical pseudorandom bit sequences (PRBS) generator is demonstrated with optical logic gate 'XNOR' and all-optical wavelength converter based on cascaded Tera-Hertz Optical Asymmetric Demultiplexer (TOADs). Its feasibility is verified by generation of return-to-zero on-off keying (RZ-OOK) 263-1 PRBS at the speed of 1 Gb/s with 10% duty radio. The high randomness of ultra-long cycle PRBS is validated by successfully passing the standard benchmark test.

  12. Second generation sequencing of the mesothelioma tumor genome.

    Directory of Open Access Journals (Sweden)

    Raphael Bueno

    Full Text Available The current paradigm for elucidating the molecular etiology of cancers relies on the interrogation of small numbers of genes, which limits the scope of investigation. Emerging second-generation massively parallel DNA sequencing technologies have enabled more precise definition of the cancer genome on a global scale. We examined the genome of a human primary malignant pleural mesothelioma (MPM tumor and matched normal tissue by using a combination of sequencing-by-synthesis and pyrosequencing methodologies to a 9.6X depth of coverage. Read density analysis uncovered significant aneuploidy and numerous rearrangements. Method-dependent informatics rules, which combined the results of different sequencing platforms, were developed to identify and validate candidate mutations of multiple types. Many more tumor-specific rearrangements than point mutations were uncovered at this depth of sequencing, resulting in novel, large-scale, inter- and intra-chromosomal deletions, inversions, and translocations. Nearly all candidate point mutations appeared to be previously unknown SNPs. Thirty tumor-specific fusions/translocations were independently validated with PCR and Sanger sequencing. Of these, 15 represented disrupted gene-encoding regions, including kinases, transcription factors, and growth factors. One large deletion in DPP10 resulted in altered transcription and expression of DPP10 transcripts in a set of 53 additional MPM tumors correlated with survival. Additionally, three point mutations were observed in the coding regions of NKX6-2, a transcription regulator, and NFRKB, a DNA-binding protein involved in modulating NFKB1. Several regions containing genes such as PCBD2 and DHFR, which are involved in growth factor signaling and nucleotide synthesis, respectively, were selectively amplified in the tumor. Second-generation sequencing uncovered all types of mutations in this MPM tumor, with DNA rearrangements representing the dominant type.

  13. Genotyping 1000 yeast strains by next-generation sequencing

    Directory of Open Access Journals (Sweden)

    Wilkening Stefan

    2013-02-01

    Full Text Available Abstract Background The throughput of next-generation sequencing machines has increased dramatically over the last few years; yet the cost and time for library preparation have not changed proportionally, thus representing the main bottleneck for sequencing large numbers of samples. Here we present an economical, high-throughput library preparation method for the Illumina platform, comprising a 96-well based method for DNA isolation for yeast cells, a low-cost DNA shearing alternative, and adapter ligation using heat inactivation of enzymes instead of bead cleanups. Results Up to 384 whole-genome libraries can be prepared from yeast cells in one week using this method, for less than 15 euros per sample. We demonstrate the robustness of this protocol by sequencing over 1000 yeast genomes at ~30x coverage. The sequence information from 768 yeast segregants derived from two divergent S. cerevisiae strains was used to generate a meiotic recombination map at unprecedented resolution. Comparisons to other datasets indicate a high conservation of recombination at a chromosome-wide scale, but differences at the local scale. Additionally, we detected a high degree of aneuploidy (3.6% by examining the sequencing coverage in these segregants. Differences in allele frequency allowed us to attribute instances of aneuploidy to gains of chromosomes during meiosis or mitosis, both of which showed a strong tendency to missegregate specific chromosomes. Conclusions Here we present a high throughput workflow to sequence genomes of large number of yeast strains at a low price. We have used this workflow to obtain recombination and aneuploidy data from hundreds of segregants, which can serve as a foundation for future studies of linkage, recombination, and chromosomal aberrations in yeast and higher eukaryotes.

  14. Interim Report: Air-Cooled Condensers for Next Generation Geothermal Power Plants Improved Binary Cycle Performance

    Energy Technology Data Exchange (ETDEWEB)

    Daniel S. Wendt; Greg L. Mines

    2010-09-01

    As geothermal resources that are more expensive to develop are utilized for power generation, there will be increased incentive to use more efficient power plants. This is expected to be the case with Enhanced Geothermal System (EGS) resources. These resources will likely require wells drilled to depths greater than encountered with hydrothermal resources, and will have the added costs for stimulation to create the subsurface reservoir. It is postulated that plants generating power from these resources will likely utilize the binary cycle technology where heat is rejected sensibly to the ambient. The consumptive use of a portion of the produced geothermal fluid for evaporative heat rejection in the conventional flash-steam conversion cycle is likely to preclude its use with EGS resources. This will be especially true in those areas where there is a high demand for finite supplies of water. Though they have no consumptive use of water, using air-cooling systems for heat rejection has disadvantages. These systems have higher capital costs, reduced power output (heat is rejected at the higher dry-bulb temperature), increased parasitics (fan power), and greater variability in power generation on both a diurnal and annual basis (larger variation in the dry-bulb temperature). This is an interim report for the task ‘Air-Cooled Condensers in Next- Generation Conversion Systems’. The work performed was specifically aimed at a plant that uses commercially available binary cycle technologies with an EGS resource. Concepts were evaluated that have the potential to increase performance, lower cost, or mitigate the adverse effects of off-design operation. The impact on both cost and performance were determined for the concepts considered, and the scenarios identified where a particular concept is best suited. Most, but not all, of the concepts evaluated are associated with the rejection of heat. This report specifically addresses three of the concepts evaluated: the use of

  15. Applications for next-generation sequencing in fish ecotoxicogenomics

    Directory of Open Access Journals (Sweden)

    Alvine C Mehinto

    2012-04-01

    Full Text Available The new technologies for next-generation sequencing and global gene expression analyses that are widely used in molecular medicine are increasingly applied to the field of fish biology. This has facilitated new directions to address research areas that could not be previously considered due to the lack of molecular information for ecologically relevant species. Over the past decade, the cost of next-generation sequencing (NGS has decreased significantly, making it possible to use non-model fish species to investigate emerging environmental issues. NGS technologies have permitted researchers to obtain large amounts of raw data in short periods of time. There have also been significant improvements in bioinformatics to assemble the sequences and annotate the genes, thus facilitating the management of these large datasets. The combination of DNA sequencing and bioinformatics has improved our abilities to design custom microarrays and study the genome and transcriptome of a wide variety of organisms. Despite the promising results obtained using these techniques in fish studies, NGS technologies are currently underused in ecotoxicogenomics and few studies have employed these methods. These issues should be addressed in order to exploit the full potential of NGS in ecotoxicological studies and expand our understanding of the biology of non-model organisms.

  16. Probing the SELEX process with next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Tatjana Schütze

    Full Text Available BACKGROUND: SELEX is an iterative process in which highly diverse synthetic nucleic acid libraries are selected over many rounds to finally identify aptamers with desired properties. However, little is understood as how binders are enriched during the selection course. Next-generation sequencing offers the opportunity to open the black box and observe a large part of the population dynamics during the selection process. METHODOLOGY: We have performed a semi-automated SELEX procedure on the model target streptavidin starting with a synthetic DNA oligonucleotide library and compared results obtained by the conventional analysis via cloning and Sanger sequencing with next-generation sequencing. In order to follow the population dynamics during the selection, pools from all selection rounds were barcoded and sequenced in parallel. CONCLUSIONS: High affinity aptamers can be readily identified simply by copy number enrichment in the first selection rounds. Based on our results, we suggest a new selection scheme that avoids a high number of iterative selection rounds while reducing time, PCR bias, and artifacts.

  17. Authentication of Herbal Supplements Using Next-Generation Sequencing

    OpenAIRE

    Ivanova, Natalia V.; Kuzmina, Maria L.; Thomas W A Braukmann; Borisenko, Alex V.; Zakharov, Evgeny V.

    2016-01-01

    Background DNA-based testing has been gaining acceptance as a tool for authentication of a wide range of food products; however, its applicability for testing of herbal supplements remains contentious. Methods We utilized Sanger and Next-Generation Sequencing (NGS) for taxonomic authentication of fifteen herbal supplements representing three different producers from five medicinal plants: Echinacea purpurea, Valeriana officinalis, Ginkgo biloba, Hypericum perforatum and Trigonella foenum-grae...

  18. Genotype and SNP calling from next-generation sequencing data

    DEFF Research Database (Denmark)

    Nielsen, Rasmus; Paul, Joshua S.; Albrechtsen, Anders

    2011-01-01

    Meaningful analysis of next-generation sequencing (NGS) data, which are produced extensively by genetics and genomics studies, relies crucially on the accurate calling of SNPs and genotypes. Recently developed statistical methods both improve and quantify the considerable uncertainty associated w...... with genotype calling, and will especially benefit the growing number of studies using low- to medium-coverage data. We review these methods and provide a guide for their use in NGS studies....

  19. Transcriptome sequencing of the Microarray Quality Control (MAQC RNA reference samples using next generation sequencing

    Directory of Open Access Journals (Sweden)

    Thierry-Mieg Danielle

    2009-06-01

    Full Text Available Abstract Background Transcriptome sequencing using next-generation sequencing platforms will soon be competing with DNA microarray technologies for global gene expression analysis. As a preliminary evaluation of these promising technologies, we performed deep sequencing of cDNA synthesized from the Microarray Quality Control (MAQC reference RNA samples using Roche's 454 Genome Sequencer FLX. Results We generated more that 3.6 million sequence reads of average length 250 bp for the MAQC A and B samples and introduced a data analysis pipeline for translating cDNA read counts into gene expression levels. Using BLAST, 90% of the reads mapped to the human genome and 64% of the reads mapped to the RefSeq database of well annotated genes with e-values ≤ 10-20. We measured gene expression levels in the A and B samples by counting the numbers of reads that mapped to individual RefSeq genes in multiple sequencing runs to evaluate the MAQC quality metrics for reproducibility, sensitivity, specificity, and accuracy and compared the results with DNA microarrays and Quantitative RT-PCR (QRTPCR from the MAQC studies. In addition, 88% of the reads were successfully aligned directly to the human genome using the AceView alignment programs with an average 90% sequence similarity to identify 137,899 unique exon junctions, including 22,193 new exon junctions not yet contained in the RefSeq database. Conclusion Using the MAQC metrics for evaluating the performance of gene expression platforms, the ExpressSeq results for gene expression levels showed excellent reproducibility, sensitivity, and specificity that improved systematically with increasing shotgun sequencing depth, and quantitative accuracy that was comparable to DNA microarrays and QRTPCR. In addition, a careful mapping of the reads to the genome using the AceView alignment programs shed new light on the complexity of the human transcriptome including the discovery of thousands of new splice variants.

  20. All-Optical Quantum Random Bit Generation from Intrinsically Binary Phase of Parametric Oscillators

    CERN Document Server

    Marandi, Alireza; Vodopyanov, Konstantin L; Byer, Robert L

    2012-01-01

    True random number generators (RNGs) are desirable for applications ranging from cryptogra- phy to computer simulations. Quantum phenomena prove to be attractive for physical RNGs due to their fundamental randomness and immunity to attack [1]- [5]. Optical parametric down conversion is an essential element in most quantum optical experiments including optical squeezing [9], and generation of entangled photons [10]. In an optical parametric oscillator (OPO), photons generated through spontaneous down conversion of the pump initiate the oscillation in the absence of other inputs [11, 12]. This quantum process is the dominant effect during the oscillation build-up, leading to selection of one of the two possible phase states above threshold in a degenerate OPO [13]. Building on this, we demonstrate a novel all-optical quantum RNG in which the photodetection is not a part of the random process, and no post processing is required for the generated bit sequence. We implement a synchronously pumped twin degenerate O...

  1. Next-Generation Sequencing and Genome Editing in Plant Virology

    Directory of Open Access Journals (Sweden)

    Ahmed Hadidi

    2016-08-01

    Full Text Available Next-generation sequencing (NGS has been applied to plant virology since 2009. NGS provides highly efficient, rapid, low cost DNA or RNA high-throughput sequencing of the genomes of plant viruses and viroids and of the specific small RNAs generated during the infection process. These small RNAs, which cover frequently the whole genome of the infectious agent, are 21-24 nt long and are known as vsRNAs for viruses and vd-sRNAs for viroids. NGS has been used in a number of studies in plant virology including, but not limited to, discovery of novel viruses and viroids as well as detection and identification of those pathogens already known, analysis of genome diversity and evolution, and study of pathogen epidemiology. The genome engineering editing method, clustered regularly interspaced short palindromic repeats (CRISPR-Cas9 system has been successfully used recently to engineer resistance to DNA geminiviruses (family, Geminiviridae by targeting different viral genome sequences in infected Nicotiana benthamiana or Arabidopsis plants. The DNA viruses targeted include tomato yellow leaf curl virus and merremia mosaic virus (begomovirus; beet curly top virus and beet severe curly top virus (curtovirus; and bean yellow dwarf virus (mastrevirus. The technique has also been used against the RNA viruses zucchini yellow mosaic virus, papaya ringspot virus and turnip mosaic virus (potyvirus and cucumber vein yellowing virus (ipomovirus, family, Potyviridae by targeting the translation initiation genes eIF4E in cucumber or Arabidopsis plants. From these recent advances of major importance, it is expected that NGS and CRISPR-Cas technologies will play a significant role in the very near future in advancing the field of plant virology and connecting it with other related fields of biology.Keywords: Next-generation sequencing, NGS, plant virology, plant viruses, viroids, resistance to plant viruses by CRISPR-Cas9

  2. Applications of next-generation sequencing to phylogeography and phylogenetics.

    Science.gov (United States)

    McCormack, John E; Hird, Sarah M; Zellmer, Amanda J; Carstens, Bryan C; Brumfield, Robb T

    2013-02-01

    This is a time of unprecedented transition in DNA sequencing technologies. Next-generation sequencing (NGS) clearly holds promise for fast and cost-effective generation of multilocus sequence data for phylogeography and phylogenetics. However, the focus on non-model organisms, in addition to uncertainty about which sample preparation methods and analyses are appropriate for different research questions and evolutionary timescales, have contributed to a lag in the application of NGS to these fields. Here, we outline some of the major obstacles specific to the application of NGS to phylogeography and phylogenetics, including the focus on non-model organisms, the necessity of obtaining orthologous loci in a cost-effective manner, and the predominate use of gene trees in these fields. We describe the most promising methods of sample preparation that address these challenges. Methods that reduce the genome by restriction digest and manual size selection are most appropriate for studies at the intraspecific level, whereas methods that target specific genomic regions (i.e., target enrichment or sequence capture) have wider applicability from the population level to deep-level phylogenomics. Additionally, we give an overview of how to analyze NGS data to arrive at data sets applicable to the standard toolkit of phylogeography and phylogenetics, including initial data processing to alignment and genotype calling (both SNPs and loci involving many SNPs). Even though whole-genome sequencing is likely to become affordable rather soon, because phylogeography and phylogenetics rely on analysis of hundreds of individuals in many cases, methods that reduce the genome to a subset of loci should remain more cost-effective for some time to come.

  3. Parameter estimation for binary black holes with networks of third generation gravitational-wave detectors

    CERN Document Server

    Vitale, Salvatore

    2016-01-01

    The two binary black-hole (BBH) coalescences detected by LIGO, GW150914 and GW151226, were relatively nearby sources, with a redshift of ~0.1. As the sensitivity of Advanced LIGO and Virgo increases in the next few years, they will eventually detect heavy BBHs up to redshifts of ~1. However, these are still relatively small distances compared with the size of the Universe, or with those encountered in most areas of astrophysics. In order to study BBH during the epoch of reionization, or black holes born from population III stars, more sensitive instruments are needed. Third-generation gravitational-wave detectors, such as the Einstein Telescope or the Cosmic Explorer are already in an advanced R&D stage. These detectors will be roughly a factor of 10 more sensitive than the current generation, and be able to detect BBH mergers beyond a redshift of 20. In this paper we quantify the precision with which these new facilities will be able to estimate the parameters of stellar-mass, heavy, and intermediate-mas...

  4. Humans cannot consciously generate random numbers sequences: Polemic study.

    Science.gov (United States)

    Figurska, Małgorzata; Stańczyk, Maciej; Kulesza, Kamil

    2008-01-01

    It is widely believed, that randomness exists in Nature. In fact such an assumption underlies many scientific theories and is embedded in the foundations of quantum mechanics. Assuming that this hypothesis is valid one can use natural phenomena, like radioactive decay, to generate random numbers. Today, computers are capable of generating the so-called pseudorandom numbers. Such series of numbers are only seemingly random (bias in the randomness quality can be observed). Question whether people can produce random numbers, has been investigated by many scientists in the recent years. The paper "Humans can consciously generate random numbers sequences..." published recently in Medical Hypotheses made claims that were in many ways contrary to state of art; it also stated far-reaching hypotheses. So, we decided to repeat the experiments reported, with special care being taken of proper laboratory procedures. Here, we present the results and discuss possible implications in computer and other sciences.

  5. Preparation of next-generation sequencing libraries from damaged DNA.

    Science.gov (United States)

    Briggs, Adrian W; Heyn, Patricia

    2012-01-01

    Next-generation sequencing (NGS) has revolutionized ancient DNA research, especially when combined with high-throughput target enrichment methods. However, attaining high sequencing depth and accuracy from samples often remains problematic due to the damaged state of ancient DNA, in particular the extremely low copy number of ancient DNA and the abundance of uracil residues derived from cytosine deamination that lead to miscoding errors. It is therefore critical to use a highly efficient procedure for conversion of a raw DNA extract into an adaptor-ligated sequencing library, and equally important to reduce errors from uracil residues. We present a protocol for NGS library preparation that allows highly efficient conversion of DNA fragments into an adaptor-ligated form. The protocol incorporates an option to remove the vast majority of uracil miscoding lesions as part of the library preparation process. The procedure requires only two spin column purification steps and no gel purification or bead handling. Starting from an aliquot of DNA extract, a finished, highly amplified library can be generated in 5 h, or under 3 h if uracil removal is not required.

  6. Next generation sequencing and its applications in forensic genetics.

    Science.gov (United States)

    Børsting, Claus; Morling, Niels

    2015-09-01

    It has been almost a decade since the first next generation sequencing (NGS) technologies emerged and quickly changed the way genetic research is conducted. Today, full genomes are mapped and published almost weekly and with ever increasing speed and decreasing costs. NGS methods and platforms have matured during the last 10 years, and the quality of the sequences has reached a level where NGS is used in clinical diagnostics of humans. Forensic genetic laboratories have also explored NGS technologies and especially in the last year, there has been a small explosion in the number of scientific articles and presentations at conferences with forensic aspects of NGS. These contributions have demonstrated that NGS offers new possibilities for forensic genetic case work. More information may be obtained from unique samples in a single experiment by analyzing combinations of markers (STRs, SNPs, insertion/deletions, mRNA) that cannot be analyzed simultaneously with the standard PCR-CE methods used today. The true variation in core forensic STR loci has been uncovered, and previously unknown STR alleles have been discovered. The detailed sequence information may aid mixture interpretation and will increase the statistical weight of the evidence. In this review, we will give an introduction to NGS and single-molecule sequencing, and we will discuss the possible applications of NGS in forensic genetics. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  7. Sequence assembly using next generation sequencing data--challenges and solutions.

    Science.gov (United States)

    Chin, Francis Y L; Leung, Henry C M; Yiu, S M

    2014-11-01

    Sequence assembling is an important step for bioinformatics study. With the help of next generation sequencing (NGS) technology, high throughput DNA fragment (reads) can be randomly sampled from DNA or RNA molecular sequence. However, as the positions of reads being sampled are unknown, assembling process is required for combining overlapped reads to reconstruct the original DNA or RNA sequence. Compared with traditional Sanger sequencing methods, although the throughput of NGS reads increases, the read length is shorter and the error rate is higher. It introduces several problems in assembling. Moreover, paired-end reads instead of single-end reads can be sampled which contain more information. The existing assemblers cannot fully utilize this information and fails to assemble longer contigs. In this article, we will revisit the major problems of assembling NGS reads on genomic, transcriptomic, metagenomic and metatranscriptomic data. We will also describe our IDBA package for solving these problems. IDBA package has adopted several novel ideas in assembling, including using multiple k, local assembling and progressive depth removal. Compared with existence assemblers, IDBA has better performance on many simulated and real sequencing datasets.

  8. Next-generation sequencing: big data meets high performance computing.

    Science.gov (United States)

    Schmidt, Bertil; Hildebrandt, Andreas

    2017-02-02

    The progress of next-generation sequencing has a major impact on medical and genomic research. This high-throughput technology can now produce billions of short DNA or RNA fragments in excess of a few terabytes of data in a single run. This leads to massive datasets used by a wide range of applications including personalized cancer treatment and precision medicine. In addition to the hugely increased throughput, the cost of using high-throughput technologies has been dramatically decreasing. A low sequencing cost of around US$1000 per genome has now rendered large population-scale projects feasible. However, to make effective use of the produced data, the design of big data algorithms and their efficient implementation on modern high performance computing systems is required.

  9. Perspectives of Integrative Cancer Genomics in Next Generation Sequencing Era

    Directory of Open Access Journals (Sweden)

    So Mee Kwon

    2012-06-01

    Full Text Available The explosive development of genomics technologies including microarrays and next generation sequencing (NGS has provided comprehensive maps of cancer genomes, including the expression of mRNAs and microRNAs, DNA copy numbers, sequence variations, and epigenetic changes. These genome-wide profiles of the genetic aberrations could reveal the candidates for diagnostic and/or prognostic biomarkers as well as mechanistic insights into tumor development and progression. Recent efforts to establish the huge cancer genome compendium and integrative omics analyses, so-called "integromics", have extended our understanding on the cancer genome, showing its daunting complexity and heterogeneity. However, the challenges of the structured integration, sharing, and interpretation of the big omics data still remain to be resolved. Here, we review several issues raised in cancer omics data analysis, including NGS, focusing particularly on the study design and analysis strategies. This might be helpful to understand the current trends and strategies of the rapidly evolving cancer genomics research.

  10. Using next generation transcriptome sequencing to predict an ectomycorrhizal metablome.

    Energy Technology Data Exchange (ETDEWEB)

    Larsen, P. E.; Sreedasyam, A.; Trivedi, G; Podila, G. K.; Cseke, L. J.; Collart, F. R. (Biosciences Division); (On Assignment, Scientific Staffing); (Univ. of Alabama at Huntsville)

    2011-05-13

    Mycorrhizae, symbiotic interactions between soil fungi and tree roots, are ubiquitous in terrestrial ecosystems. The fungi contribute phosphorous, nitrogen and mobilized nutrients from organic matter in the soil and in return the fungus receives photosynthetically-derived carbohydrates. This union of plant and fungal metabolisms is the mycorrhizal metabolome. Understanding this symbiotic relationship at a molecular level provides important contributions to the understanding of forest ecosystems and global carbon cycling. We generated next generation short-read transcriptomic sequencing data from fully-formed ectomycorrhizae between Laccaria bicolor and aspen (Populus tremuloides) roots. The transcriptomic data was used to identify statistically significantly expressed gene models using a bootstrap-style approach, and these expressed genes were mapped to specific metabolic pathways. Integration of expressed genes that code for metabolic enzymes and the set of expressed membrane transporters generates a predictive model of the ectomycorrhizal metabolome. The generated model of mycorrhizal metabolome predicts that the specific compounds glycine, glutamate, and allantoin are synthesized by L. bicolor and that these compounds or their metabolites may be used for the benefit of aspen in exchange for the photosynthetically-derived sugars fructose and glucose. The analysis illustrates an approach to generate testable biological hypotheses to investigate the complex molecular interactions that drive ectomycorrhizal symbiosis. These models are consistent with experimental environmental data and provide insight into the molecular exchange processes for organisms in this complex ecosystem. The method used here for predicting metabolomic models of mycorrhizal systems from deep RNA sequencing data can be generalized and is broadly applicable to transcriptomic data derived from complex systems.

  11. Using next generation transcriptome sequencing to predict an ectomycorrhizal metabolome

    Directory of Open Access Journals (Sweden)

    Cseke Leland J

    2011-05-01

    Full Text Available Abstract Background Mycorrhizae, symbiotic interactions between soil fungi and tree roots, are ubiquitous in terrestrial ecosystems. The fungi contribute phosphorous, nitrogen and mobilized nutrients from organic matter in the soil and in return the fungus receives photosynthetically-derived carbohydrates. This union of plant and fungal metabolisms is the mycorrhizal metabolome. Understanding this symbiotic relationship at a molecular level provides important contributions to the understanding of forest ecosystems and global carbon cycling. Results We generated next generation short-read transcriptomic sequencing data from fully-formed ectomycorrhizae between Laccaria bicolor and aspen (Populus tremuloides roots. The transcriptomic data was used to identify statistically significantly expressed gene models using a bootstrap-style approach, and these expressed genes were mapped to specific metabolic pathways. Integration of expressed genes that code for metabolic enzymes and the set of expressed membrane transporters generates a predictive model of the ectomycorrhizal metabolome. The generated model of mycorrhizal metabolome predicts that the specific compounds glycine, glutamate, and allantoin are synthesized by L. bicolor and that these compounds or their metabolites may be used for the benefit of aspen in exchange for the photosynthetically-derived sugars fructose and glucose. Conclusions The analysis illustrates an approach to generate testable biological hypotheses to investigate the complex molecular interactions that drive ectomycorrhizal symbiosis. These models are consistent with experimental environmental data and provide insight into the molecular exchange processes for organisms in this complex ecosystem. The method used here for predicting metabolomic models of mycorrhizal systems from deep RNA sequencing data can be generalized and is broadly applicable to transcriptomic data derived from complex systems.

  12. Next-generation sequencing technology in clinical virology.

    Science.gov (United States)

    Capobianchi, M R; Giombini, E; Rozera, G

    2013-01-01

    Recent advances in nucleic acid sequencing technologies, referred to as 'next-generation' sequencing (NGS), have produced a true revolution and opened new perspectives for research and diagnostic applications, owing to the high speed and throughput of data generation. So far, NGS has been applied to metagenomics-based strategies for the discovery of novel viruses and the characterization of viral communities. Additional applications include whole viral genome sequencing, detection of viral genome variability, and the study of viral dynamics. These applications are particularly suitable for viruses such as human immunodeficiency virus, hepatitis B virus, and hepatitis C virus, whose error-prone replication machinery, combined with the high replication rate, results, in each infected individual, in the formation of many genetically related viral variants referred to as quasi-species. The viral quasi-species, in turn, represents the substrate for the selective pressure exerted by the immune system or by antiviral drugs. With traditional approaches, it is difficult to detect and quantify minority genomes present in viral quasi-species that, in fact, may have biological and clinical relevance. NGS provides, for each patient, a dataset of clonal sequences that is some order of magnitude higher than those obtained with conventional approaches. Hence, NGS is an extremely powerful tool with which to investigate previously inaccessible aspects of viral dynamics, such as the contribution of different viral reservoirs to replicating virus in the course of the natural history of the infection, co-receptor usage in minority viral populations harboured by different cell lineages, the dynamics of development of drug resistance, and the re-emergence of hidden genomes after treatment interruptions. The diagnostic application of NGS is just around the corner. © 2012 The Authors Clinical Microbiology and Infection © 2012 European Society of Clinical Microbiology and Infectious

  13. Ultrasensitive single-genome sequencing: accurate, targeted, next generation sequencing of HIV-1 RNA.

    Science.gov (United States)

    Boltz, Valerie F; Rausch, Jason; Shao, Wei; Hattori, Junko; Luke, Brian; Maldarelli, Frank; Mellors, John W; Kearney, Mary F; Coffin, John M

    2016-12-20

    Although next generation sequencing (NGS) offers the potential for studying virus populations in unprecedented depth, PCR error, amplification bias and recombination during library construction have limited its use to population sequencing and measurements of unlinked allele frequencies. Here we report a method, termed ultrasensitive Single-Genome Sequencing (uSGS), for NGS library construction and analysis that eliminates PCR errors and recombinants, and generates single-genome sequences of the same quality as the "gold-standard" of HIV-1 single-genome sequencing assay but with more than 100-fold greater depth. Primer ID tagged cDNA was synthesized from mixtures of cloned BH10 wild-type and mutant HIV-1 transcripts containing ten drug resistance mutations. First, the resultant cDNA was divided and NGS libraries were generated in parallel using two methods: uSGS and a method applying long PCR primers to attach the NGS adaptors (LP-PCR-1). Second, cDNA was divided and NGS libraries were generated in parallel comparing 3 methods: uSGS and 2 methods adapted from more recent reports using variations of the long PCR primers to attach the adaptors (LP-PCR-2 and LP-PCR-3). Consistently, the uSGS method amplified a greater proportion of cDNAs, averaging 30% compared to 13% for LP-PCR-1, 21% for LP-PCR-2 and 14% for LP-PCR-3. Most importantly, when the uSGS sequences were binned according to their primer IDs, 94% of the bins did not contain PCR recombinant sequences versus only 55, 75 and 65% for LP-PCR-1, 2 and 3, respectively. Finally, when uSGS was applied to plasma samples from HIV-1 infected donors, both frequent and rare variants were detected in each sample and neighbor-joining trees revealed clusters of genomes driven by the linkage of these mutations, showing the lack of PCR recombinants in the datasets. The uSGS assay can be used for accurate detection of rare variants and for identifying linkage of rare alleles associated with HIV-1 drug resistance. In addition

  14. Use of next-generation sequencing in oral cavity cancer

    DEFF Research Database (Denmark)

    Tabatabaeifar, Siavosh; Kruse, Torben A; Thomassen, Mads

    Background: Oral cavity cancer is a subgroup of head and neck cancer which is the world’s 6th most common cancer form. Oral squamous cell carcinomas (OSCC) constitute almost all oral cavity cancers, and OSCC are primarily attributed by excessive alcohol consumption and tobacco exposure...... of tumour cells exists. Conclusions: Use of next generation sequencing in oral cavity cancer can give valuable insight into the biology of the disease. By investigating intra tumour heterogeneity we see that the different tumour specimens in each patient are quite homogenous, but evidence of heterogeneous...

  15. Next-Generation Sequencing-Based Molecular Diagnosis of Choroideremia

    Directory of Open Access Journals (Sweden)

    Kayo Shimizu

    2015-07-01

    Full Text Available We screened patients with choroideremia using next-generation sequencing (NGS and identified a novel mutation and a known mutation in the CHM gene. One patient presented an atypical fundus appearance for choroideremia. Another patient presented macular hole retinal detachment in the left eye. The present case series shows the utility of NGS-based screening in patients with choroideremia. In addition, the presence of macular hole in 1 of the 2 patients, together with a previous report, indicated the susceptibility of patients with choroideremia to macular hole.

  16. Mapping sensorimotor sequences to word sequences: a connectionist model of language acquisition and sentence generation.

    Science.gov (United States)

    Takac, Martin; Benuskova, Lubica; Knott, Alistair

    2012-11-01

    In this article we present a neural network model of sentence generation. The network has both technical and conceptual innovations. Its main technical novelty is in its semantic representations: the messages which form the input to the network are structured as sequences, so that message elements are delivered to the network one at a time. Rather than learning to linearise a static semantic representation as a sequence of words, our network rehearses a sequence of semantic signals, and learns to generate words from selected signals. Conceptually, the network's use of rehearsed sequences of semantic signals is motivated by work in embodied cognition, which posits that the structure of semantic representations has its origin in the serial structure of sensorimotor processing. The rich sequential structure of the network's semantic inputs also allows it to incorporate certain Chomskyan ideas about innate syntactic knowledge and parameter-setting, as well as a more empiricist account of the acquisition of idiomatic syntactic constructions. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Change in the orbital period of a binary system due to dynamical tides for main-sequence stars

    Science.gov (United States)

    Chernov, S. V.

    2017-03-01

    We investigate the change in the orbital period of a binary system due to dynamical tides by taking into account the evolution of a main-sequence star. Three stars with masses of one, one and a half, and two solar masses are considered. A star of one solar mass at lifetimes t = 4.57 × 109 yr closely corresponds to our Sun. We show that a planet of one Jupiter mass revolving around a star of one solar mass will fall onto the star in the main-sequence lifetime of the star due to dynamical tides if the initial orbital period of the planet is less than P orb ≈ 2.8 days. Planets of one Jupiter mass with an orbital period P orb ≈ 2 days or shorter will fall onto a star of one and a half and two solar masses in the mainsequence lifetime of the star.

  18. Targeted sequencing of cancer-related genes in colorectal cancer using next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Sae-Won Han

    Full Text Available Recent advance in sequencing technology has enabled comprehensive profiling of genetic alterations in cancer. We have established a targeted sequencing platform using next-generation sequencing (NGS technology for clinical use, which can provide mutation and copy number variation data. NGS was performed with paired-end library enriched with exons of 183 cancer-related genes. Normal and tumor tissue pairs of 60 colorectal adenocarcinomas were used to test feasibility. Somatic mutation and copy number alteration were analyzed. A total of 526 somatic non-synonymous sequence variations were found in 113 genes. Among these, 278 single nucleotide variations were 232 different somatic point mutations. 216 SNV were 79 known single nucleotide polymorphisms in the dbSNP. 32 indels were 28 different indel mutations. Median number of mutated gene per tumor was 4 (range 0-23. Copy number gain (>X2 fold was found in 65 genes in 40 patients, whereas copy number loss (sequencing platform using NGS technology is feasible for clinical use and provides comprehensive genetic alteration data.

  19. Improvement of SNR with Chaotic Spreading Sequences for CDMA

    CERN Document Server

    Umeno, K; Umeno, Ken; Kitayama, Ken-ichi

    1999-01-01

    We show that chaotic spreading sequences generated by ergodic mappings of Chebyshev orthogonal polynomials have better correlation properties for CDMA(code division multiple access) than the optimal binary sequences (Gold sequences) in the sense of ensemble average.

  20. Application of next-generation sequencing technologies in virology.

    Science.gov (United States)

    Radford, Alan D; Chapman, David; Dixon, Linda; Chantrey, Julian; Darby, Alistair C; Hall, Neil

    2012-09-01

    The progress of science is punctuated by the advent of revolutionary technologies that provide new ways and scales to formulate scientific questions and advance knowledge. Following on from electron microscopy, cell culture and PCR, next-generation sequencing is one of these methodologies that is now changing the way that we understand viruses, particularly in the areas of genome sequencing, evolution, ecology, discovery and transcriptomics. Possibilities for these methodologies are only limited by our scientific imagination and, to some extent, by their cost, which has restricted their use to relatively small numbers of samples. Challenges remain, including the storage and analysis of the large amounts of data generated. As the chemistries employed mature, costs will decrease. In addition, improved methods for analysis will become available, opening yet further applications in virology including routine diagnostic work on individuals, and new understanding of the interaction between viral and host transcriptomes. An exciting era of viral exploration has begun, and will set us new challenges to understand the role of newly discovered viral diversity in both disease and health.

  1. Next-Generation Sequencing and Genome Editing in Plant Virology.

    Science.gov (United States)

    Hadidi, Ahmed; Flores, Ricardo; Candresse, Thierry; Barba, Marina

    2016-01-01

    Next-generation sequencing (NGS) has been applied to plant virology since 2009. NGS provides highly efficient, rapid, low cost DNA, or RNA high-throughput sequencing of the genomes of plant viruses and viroids and of the specific small RNAs generated during the infection process. These small RNAs, which cover frequently the whole genome of the infectious agent, are 21-24 nt long and are known as vsRNAs for viruses and vd-sRNAs for viroids. NGS has been used in a number of studies in plant virology including, but not limited to, discovery of novel viruses and viroids as well as detection and identification of those pathogens already known, analysis of genome diversity and evolution, and study of pathogen epidemiology. The genome engineering editing method, clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been successfully used recently to engineer resistance to DNA geminiviruses (family, Geminiviridae) by targeting different viral genome sequences in infected Nicotiana benthamiana or Arabidopsis plants. The DNA viruses targeted include tomato yellow leaf curl virus and merremia mosaic virus (begomovirus); beet curly top virus and beet severe curly top virus (curtovirus); and bean yellow dwarf virus (mastrevirus). The technique has also been used against the RNA viruses zucchini yellow mosaic virus, papaya ringspot virus and turnip mosaic virus (potyvirus) and cucumber vein yellowing virus (ipomovirus, family, Potyviridae) by targeting the translation initiation genes eIF4E in cucumber or Arabidopsis plants. From these recent advances of major importance, it is expected that NGS and CRISPR-Cas technologies will play a significant role in the very near future in advancing the field of plant virology and connecting it with other related fields of biology.

  2. Next-Generation Sequencing and Genome Editing in Plant Virology

    Science.gov (United States)

    Hadidi, Ahmed; Flores, Ricardo; Candresse, Thierry; Barba, Marina

    2016-01-01

    Next-generation sequencing (NGS) has been applied to plant virology since 2009. NGS provides highly efficient, rapid, low cost DNA, or RNA high-throughput sequencing of the genomes of plant viruses and viroids and of the specific small RNAs generated during the infection process. These small RNAs, which cover frequently the whole genome of the infectious agent, are 21–24 nt long and are known as vsRNAs for viruses and vd-sRNAs for viroids. NGS has been used in a number of studies in plant virology including, but not limited to, discovery of novel viruses and viroids as well as detection and identification of those pathogens already known, analysis of genome diversity and evolution, and study of pathogen epidemiology. The genome engineering editing method, clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been successfully used recently to engineer resistance to DNA geminiviruses (family, Geminiviridae) by targeting different viral genome sequences in infected Nicotiana benthamiana or Arabidopsis plants. The DNA viruses targeted include tomato yellow leaf curl virus and merremia mosaic virus (begomovirus); beet curly top virus and beet severe curly top virus (curtovirus); and bean yellow dwarf virus (mastrevirus). The technique has also been used against the RNA viruses zucchini yellow mosaic virus, papaya ringspot virus and turnip mosaic virus (potyvirus) and cucumber vein yellowing virus (ipomovirus, family, Potyviridae) by targeting the translation initiation genes eIF4E in cucumber or Arabidopsis plants. From these recent advances of major importance, it is expected that NGS and CRISPR-Cas technologies will play a significant role in the very near future in advancing the field of plant virology and connecting it with other related fields of biology. PMID:27617007

  3. Discovery of posttranscriptional regulatory RNAs using next generation sequencing technologies.

    Science.gov (United States)

    Gelderman, Grant; Contreras, Lydia M

    2013-01-01

    Next generation sequencing (NGS) has revolutionized the way by which we engineer metabolism by radically altering the path to genome-wide inquiries. This is due to the fact that NGS approaches offer several powerful advantages over traditional methods that include the ability to fully sequence hundreds to thousands of genes in a single experiment and simultaneously detect homozygous and heterozygous deletions, alterations in gene copy number, insertions, translocations, and exome-wide substitutions that include "hot-spot mutations." This chapter describes the use of these technologies as a sequencing technique for transcriptome analysis and discovery of regulatory RNA elements in the context of three main platforms: Illumina HiSeq, 454 pyrosequencing, and SOLiD sequencing. Specifically, this chapter focuses on the use of Illumina HiSeq, since it is the most widely used platform for RNA discovery and transcriptome analysis. Regulatory RNAs have now been found in all branches of life. In bacteria, noncoding small RNAs (sRNAs) are involved in highly sophisticated regulatory circuits that include quorum sensing, carbon metabolism, stress responses, and virulence (Gorke and Vogel, Gene Dev 22:2914-2925, 2008; Gottesman, Trends Genet 21:399-404, 2005; Romby et al., Curr Opin Microbiol 9:229-236, 2006). Further characterization of the underlying regulation of gene expression remains poorly understood given that it is estimated that over 60% of all predicted genes remain hypothetical and the 5' and 3' untranslated regions are unknown for more than 90% of the genes (Siegel et al., Trends Parasitol 27:434-441, 2011). Importantly, manipulation of the posttranscriptional regulation that occurs at the level of RNA stability and export, trans-splicing, polyadenylation, protein translation, and protein stability via untranslated regions (Clayton, EMBO J 21:1881-1888, 2002; Haile and Papadopoulou, Curr Opin Microbiol 10:569-577, 2007) could be highly beneficial to metabolic

  4. QTrim : a novel tool for the quality trimming of sequence reads generated using the Roche/454 sequencing platform

    OpenAIRE

    Shrestha, Ram; Lubinsky, Baruch; Bansode, Vijay B; Moinz, Mónica B. J.; McCormack, Grace P.; Travers, Simon A

    2014-01-01

    Background\\ud Many high throughput sequencing (HTS) approaches, such as the Roche/454 platform, produce sequences in which the quality of the sequence (as measured by a Phred-like quality scores) decreases linearly across a sequence read. Undertaking quality trimming of this data is essential to enable confidence in the results of subsequent downstream analysis. Here, we have developed a novel, highly sensitive and accurate approach (QTrim) for the quality trimming of sequence reads generated...

  5. PAFFT: A new homology search algorithm for third-generation sequencers.

    Science.gov (United States)

    Misawa, Kazuharu; Ootsuki, Ryo

    2015-11-01

    DNA sequencers that can conduct real-time sequencing from a single polymerase molecule are known as third-generation sequencers. Third-generation sequencers enable sequencing of reads that are several kilobases long. However, the raw data generated from third-generation sequencers are known to be error-prone. Because of sequencing errors, it is difficult to identify which genes are homologous to the reads obtained using third-generation sequencers. In this study, a new method for homology search algorithm, PAFFT, is developed. This method is the extension of the MAFFT algorithm which was used for multiple alignments. PAFFT detects global homology rather than local homology so that homologous regions can be detected even when the error rate of sequencing is high. PAFFT will boost application of third-generation sequencers. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Characterization of sequence-specific errors in various next-generation sequencing systems.

    Science.gov (United States)

    Shin, Sunguk; Park, Joonhong

    2016-03-01

    Next-generation sequencing (NGS) is a popular method for assessing the molecular diversity of microbial communities without cultivation, for identifying polymorphisms in populations, and for comparing genomes and transcriptomes. However, sequence-specific errors (SSEs) by NGS systems can result in genome mis-assembly, overestimation of diversity in microbial community analyses, and false polymorphism discovery. SSEs can be particularly problematic due to rich microbial biodiversity and genomes containing frequent repeats. In this study, SSEs in public data from all popular NGS systems were discovered using a Markov chain model and hotspots for sequence errors were identified. Deletion errors were frequently preceded by homopolymers in non-Illumina NGS systems, such as GS FLX+. Substitution errors were often related to high GC contents and long G/C homopolymers in Illumina sequencing systems such as HiSeq. After removal of long G/C homopolymers in HiSeq, the average lengths of contigs and average SNP quality increased. SSEs were selectively removed from our mock community data by quality filtering, and a bias against specific microbes was identified. Our findings provide a scientific basis for filtering poor-quality reads, correcting deletion errors, preventing genome mis-assembly, and accurately assessing microbial community compositions and polymorphisms.

  7. Absolute properties of the main-sequence eclipsing binary FM Leo

    CERN Document Server

    Ratajczak, M; Schwarzenberg-Czerny, A; Dimitrov, W; Konacki, M; Helminiak, K G; Bartczak, P; Fagas, M; Kaminski, K; Kankiewicz, P; Borczyk, W; Rozek, A

    2009-01-01

    First spectroscopic and new photometric observations of the eclipsing binary FM Leo are presented. The main aims were to determine orbital and stellar parameters of two components and their evolutionary stage. First spectroscopic observations of the system were obtained with DDO and PST spectrographs. The results of the orbital solution from radial velocity curves are combined with those derived from the light-curve analysis (ASAS-3 photometry and supplementary observations of eclipses with 1 m and 0.35 m telescopes) to derive orbital and stellar parameters. JKTEBOP, Wilson-Devinney binary modelling codes and a two-dimensional cross-correlation (TODCOR) method were applied for the analysis. We find the masses to be M_1 = 1.318 $\\pm$ 0.007 and M_2 = 1.287 $\\pm$ 0.007 M_sun, the radii to be R_1 = 1.648 $\\pm$ 0.043 and R_2 = 1.511 $\\pm$ 0.049 R_sun for primary and secondary stars, respectively. The evolutionary stage of the system is briefly discussed by comparing physical parameters with current stellar evoluti...

  8. Next generation sequencing for characterizing biodiversity: promises and challenges.

    Science.gov (United States)

    Pompanon, François; Samadi, Sarah

    2015-04-01

    DNA barcoding approaches are used to describe biodiversity by analysing specimens or environmental samples in taxonomic, phylogenetic and ecological studies. While sharing data among these disciplines would be highly valuable, this remains difficult because of contradictory requirements. The properties making a DNA barcode efficient for specimen identification or species delimitation are hardly reconcilable with those required for a powerful analysis of degraded DNA from environmental samples. The use of next generation sequencing methods open up the way towards the development of new markers (e.g., multilocus barcodes) that would overcome such limitations. However, several challenges should be taken up for coordinating actions at the interface between taxonomy, ecology, molecular biology and bioinformatics in order to develop methods and protocols compatible with both taxonomic and ecological studies.

  9. Application of next-generation sequencing technology in forensic science.

    Science.gov (United States)

    Yang, Yaran; Xie, Bingbing; Yan, Jiangwei

    2014-10-01

    Next-generation sequencing (NGS) technology, with its high-throughput capacity and low cost, has developed rapidly in recent years and become an important analytical tool for many genomics researchers. New opportunities in the research domain of the forensic studies emerge by harnessing the power of NGS technology, which can be applied to simultaneously analyzing multiple loci of forensic interest in different genetic contexts, such as autosomes, mitochondrial and sex chromosomes. Furthermore, NGS technology can also have potential applications in many other aspects of research. These include DNA database construction, ancestry and phenotypic inference, monozygotic twin studies, body fluid and species identification, and forensic animal, plant and microbiological analyses. Here we review the application of NGS technology in the field of forensic science with the aim of providing a reference for future forensics studies and practice.

  10. Unrevealed mosaicism in the next-generation sequencing era.

    Science.gov (United States)

    Gajecka, Marzena

    2016-04-01

    Mosaicism refers to the presence in an individual of normal and abnormal cells that are genotypically distinct and are derived from a single zygote. The incidence of mosaicism events in the human body is underestimated as the genotypes in the mosaic ratio, especially in the low-grade mosaicism, stay unrevealed. This review summarizes various research outcomes and diagnostic questions in relation to different types of mosaicism. The impact of both tested biological material and applied method on the mosaicism detection rate is especially highlighted. As next-generation sequencing technologies constitute a promising methodological solution in mosaicism detection in the coming years, revisions in current diagnostic protocols are necessary to increase the detection rate of the unrevealed mosaicism events. Since mosaicism identification is a complex process, numerous examples of multistep mosaicism investigations are presented and discussed.

  11. Application of Next-generation Sequencing Technology in Forensic Science

    Directory of Open Access Journals (Sweden)

    Yaran Yang

    2014-10-01

    Full Text Available Next-generation sequencing (NGS technology, with its high-throughput capacity and low cost, has developed rapidly in recent years and become an important analytical tool for many genomics researchers. New opportunities in the research domain of the forensic studies emerge by harnessing the power of NGS technology, which can be applied to simultaneously analyzing multiple loci of forensic interest in different genetic contexts, such as autosomes, mitochondrial and sex chromosomes. Furthermore, NGS technology can also have potential applications in many other aspects of research. These include DNA database construction, ancestry and phenotypic inference, monozygotic twin studies, body fluid and species identification, and forensic animal, plant and microbiological analyses. Here we review the application of NGS technology in the field of forensic science with the aim of providing a reference for future forensics studies and practice.

  12. Applications of Next-generation Sequencing in Systemic Autoimmune Diseases

    Directory of Open Access Journals (Sweden)

    Yiyangzi Ma

    2015-08-01

    Full Text Available Systemic autoimmune diseases are a group of heterogeneous disorders caused by both genetic and environmental factors. Although numerous causal genes have been identified by genome-wide association studies (GWAS, these susceptibility genes are correlated to a relatively low disease risk, indicating that environmental factors also play an important role in the pathogenesis of disease. The intestinal microbiome, as the main symbiotic ecosystem between the host and host-associated microorganisms, has been demonstrated to regulate the development of the body’s immune system and is likely related to genetic mutations in systemic autoimmune diseases. Next-generation sequencing (NGS technology, with high-throughput capacity and accuracy, provides a powerful tool to discover genomic mutations, abnormal transcription and intestinal microbiome identification for autoimmune diseases. In this review, we briefly outlined the applications of NGS in systemic autoimmune diseases. This review may provide a reference for future studies in the pathogenesis of systemic autoimmune diseases.

  13. Applications of Next-generation Sequencing in Systemic Autoimmune Diseases

    Institute of Scientific and Technical Information of China (English)

    Yiyangzi Ma; Na Shi; Mengtao Li; Fei Chen; Haitao Niu

    2015-01-01

    Systemic autoimmune diseases are a group of heterogeneous disorders caused by both genetic and environmental factors. Although numerous causal genes have been identified by genome-wide association studies (GWAS), these susceptibility genes are correlated to a relatively low disease risk, indicating that environmental factors also play an important role in the pathogen-esis of disease. The intestinal microbiome, as the main symbiotic ecosystem between the host and host-associated microorganisms, has been demonstrated to regulate the development of the body’s immune system and is likely related to genetic mutations in systemic autoimmune diseases. Next-generation sequencing (NGS) technology, with high-throughput capacity and accuracy, provides a powerful tool to discover genomic mutations, abnormal transcription and intestinal microbiome identification for autoimmune diseases. In this review, we briefly outlined the applications of NGS in systemic autoimmune diseases. This review may provide a reference for future studies in the pathogenesis of systemic autoimmune diseases.

  14. Field guide to next-generation DNA sequencers.

    Science.gov (United States)

    Glenn, Travis C

    2011-09-01

    The diversity of available 2(nd) and 3(rd) generation DNA sequencing platforms is increasing rapidly. Costs for these systems range from $10/Mb for 454 and some Ion Torrent chips). In terms of cost per nonmultiplexed sample and instrument run time, the Pacific Biosciences and Ion Torrent platforms excel, with the 454 GS Junior and Illumina MiSeq also notable in this regard. All platforms allow multiplexing of samples, but details of library preparation, experimental design and data analysis can constrain the options. The wide range of characteristics among available platforms provides opportunities both to conduct groundbreaking studies and to waste money on scales that were previously infeasible. Thus, careful thought about the desired characteristics of these systems is warranted before purchasing or using any of them. Updated information from this guide will be maintained at: http://dna.uga.edu/ and http://tomato.biol.trinity.edu/blog/. © 2011 Blackwell Publishing Ltd.

  15. Application of Next-generation Sequencing Technology in Forensic Science

    Institute of Scientific and Technical Information of China (English)

    Yaran Yang; Bingbing Xie; Jiangwei Yan

    2014-01-01

    Next-generation sequencing (NGS) technology, with its high-throughput capacity and low cost, has developed rapidly in recent years and become an important analytical tool for many genomics researchers. New opportunities in the research domain of the forensic studies emerge by harnessing the power of NGS technology, which can be applied to simultaneously analyzing multi-ple loci of forensic interest in different genetic contexts, such as autosomes, mitochondrial and sex chromosomes. Furthermore, NGS technology can also have potential applications in many other aspects of research. These include DNA database construction, ancestry and phenotypic inference, monozygotic twin studies, body fluid and species identification, and forensic animal, plant and microbiological analyses. Here we review the application of NGS technology in the field of forensic science with the aim of providing a reference for future forensics studies and practice.

  16. Anomalous statistics of aftershock sequences generated by supershear ruptures

    Directory of Open Access Journals (Sweden)

    Pathikrit Bhattacharya

    2012-05-01

    Full Text Available Most earthquake ruptures propagate with speeds smaller than the Rayleigh wave velocity of the medium. These are called sub- Rayleigh ruptures. However, under suitable conditions, segments of otherwise sub- Rayleigh seismogenic ruptures can occasionally accelerate to speeds higher than the local shear wave velocity, giving rise to so-called supershear ruptures. The occurrence of supershear ruptures is usually associated with a locally higher value of pre-stress on the fault segment compared to the sub-Rayleigh segments of the same fault. Additionally, shear stress changes generated by the supershear rupture are radiated out unattenuated to distances comparable to the depth of rupture instead of rapidly decaying at much smaller distances from the rupture. This leads to aftershocks being distributed away from the fault on the supershear segment. This study attempts to verify whether these pre- and postseismic stress conditions and the resultant spatial aftershock distributions lead to discernible features in the statistical properties of the aftershock sequences of the earthquakes known to be associated with supershear ruptures. We analyze the Gutenberg-Richter scaling, the modified Omori law and Båth’s law for the aftershock sequences of two supershear mainshocks: the 1979 MW 6.5 Imperial Valley (California and 2002 MW 7.9 Denali (Alaska earthquakes. We observe that the b-value is always higher in the supershear zone than the rest of the sequence. We also observe that there is no systematic trend in the exponent of the modified Omori law when comparing the aftershocks in the supershear zone with the rest of the aftershocks. We argue that the b-value anomaly can be explained in terms of the off-fault distribution of aftershocks around the supershear segment of the rupture.

  17. The impact of next-generation sequencing on genomics

    Institute of Scientific and Technical Information of China (English)

    Jun Zhang; Rod Chiodini; Ahmed Badr; Genfa Zhang

    2011-01-01

    This article reviews basic concepts,general applications,and the potential impact of next-generation sequencing(NGS)technologies on genomics,with particular reference to currently available and possible future platforms and bioinformatics.NGS technologies have demonstrated the capacity to sequence DNA at unprecedented speed,thereby enabling previously unimaginable scientific achievements and novel biological applications.But,the massive data produced by NGS also presents a significant challenge for data storage,analyses,and management solutions.Advanced bioinformatic tools are essential for the successful application of NGS technology.As evidenced throughout this review,NGS technologies will have a striking impact on genomic research and the entire biological field.With its ability to tackle the unsolved challenges unconquered by previous genomic technologies,NGS is likely to unravel the complexity of the human genome in terms of genetic variations,some of which may be confined to susceptible loci for some common human conditions.The impact of NGS technologies on genomics will be far reaching and likely change the field for years to come.

  18. Measuring the mass of a pre-main sequence binary star through the orbit of TWA5A

    Energy Technology Data Exchange (ETDEWEB)

    Konopacky, Q; Ghez, A; Duchene, G; McCabe, C; Macintosh, B

    2007-01-18

    We present the results of a five year monitoring campaign of the close binary TWA 5Aab in the TW Hydrae association, using speckle and adaptive optics on the W.M. Keck 10 m telescopes. These measurements were taken as part of our ongoing monitoring of pre-main sequence (PMS) binaries in an effort to increase the number of dynamically determined PMS masses and thereby calibrate the theoretical PMS evolutionary tracks. Our observations have allowed us to obtain the first determination of this system's astrometric orbit. We find an orbital period of 5.94 {+-} 0.09 years and a semi-major axis of 0.''066 {+-} 0.''005. Combining these results with a kinematic distance, we calculate a total mass of 0.71 {+-} 0.14 M{sub {circle_dot}} (D/44 pc){sup 3}. for this system. This mass measurement, as well as the estimated age of this system, are consistent to within 2{sigma} of all theoretical models considered. In this analysis, we properly account for correlated uncertainties, and show that while these correlations are generally ignored, they increase the formal uncertainties by up to a factor of five and therefore are important to incorporate. With only a few more years of observation, this type of measurement will allow the theoretical models to be distinguished.

  19. Two new SB2 binaries with main sequence B-type pulsators in the Kepler field

    CERN Document Server

    Pápics, P I; Aerts, C; Briquet, M; Marcos-Arenal, P; Beck, P G; Uytterhoeven, K; Hage, A Triviño; Southworth, J; Clubb, K I; Bloemen, S; Degroote, P; Jackiewicz, J; McKeever, J; Van Winckel, H; Niemczura, E; Gameiro, J F; Debosscher, J

    2013-01-01

    Context: OB stars are important in the chemistry and evolution of the Universe, but the sample of targets well understood from an asteroseismological point of view is still too limited to provide feedback on the current evolutionary models. Our study extends this sample with two spectroscopic binary systems. AIMS. Our goal is to provide orbital solutions, fundamental parameters and abundances from disentangled high-resolution high signal-to-noise spectra, as well as to analyse and interpret the variations in the Kepler light curve of these carefully selected targets. This way we continue our efforts to map the instability strips of beta Cep and SPB stars using the combination of high-resolution ground-based spectroscopy and uninterrupted space-based photometry. Methods: We fit Keplerian orbits to radial velocities measured from selected absorption lines of high-resolution spectroscopy using synthetic composite spectra to obtain orbital solutions. We use revised masks to obtain optimal light curves from the or...

  20. Human identification by lice: A Next Generation Sequencing challenge.

    Science.gov (United States)

    Pilli, Elena; Agostino, Alessandro; Vergani, Debora; Salata, Elena; Ciuna, Ignazio; Berti, Andrea; Caramelli, David; Lambiase, Simonetta

    2016-09-01

    Rapid and progressive advances in molecular biology techniques and the advent of Next Generation Sequencing (NGS) have opened new possibilities for analyses also in the identification of entomological matrixes. Insects and other arthropods are widespread in nature and those found at a crime scene can provide a useful contribution to forensic investigations. Entomological evidence is used by experts to define the postmortem interval (PMI), which is essentially based on morphological recognition of the insect and an estimation of its insect life cycle stage. However, molecular genotyping methods can also provide an important support for forensic entomological investigations when the identification of species or human genetic material is required. This case study concerns a collection of insects found in the house of a woman who died from unknown causes. Initially the insects were identified morphologically as belonging to the Pediculidae family, and then, human DNA was extracted and analyzed from their gastrointestinal tract. The application of the latest generation forensic DNA assays, such as the Quantifiler(®) Trio DNA Quantification Kit and the HID-Ion AmpliSeq™ Identity Panel (Applied Biosystems(®)), individuated the presence of human DNA in the samples and determined the genetic profile.

  1. The Great Escape III: Placing post-main-sequence evolution of planetary and binary systems in a Galactic context

    CERN Document Server

    Veras, Dimitri; Wyatt, Mark C; Tout, Christopher A

    2013-01-01

    Our improving understanding of the life cycle of planetary systems prompts investigations of the role of the Galactic environment before, during and after Asymptotic Giant Branch (AGB) stellar evolution. Here, we investigate the interplay between stellar mass loss, Galactic tidal perturbations, and stellar flybys for evolving stars which host one planet, smaller body or stellar binary companion and reside in the Milky Way's bulge or disc. We find that the potential evolutionary pathways from a main sequence (MS) to a white dwarf (WD) planetary system are a strong function of Galactocentric distance only with respect to the prevalence of stellar flybys. Planetary ejection and collision with the parent star should be more common towards the bulge. At a given location anywhere in the Galaxy, if the mass loss is adiabatic, then the secondary is likely to avoid close flybys during AGB evolution, and cannot eventually escape the resulting WD because of Galactic tides alone. Partly because AGB mass loss will shrink ...

  2. Facial Expression Recognition from Video Sequences Based on Spatial-Temporal Motion Local Binary Pattern and Gabor Multiorientation Fusion Histogram

    Directory of Open Access Journals (Sweden)

    Lei Zhao

    2017-01-01

    Full Text Available This paper proposes novel framework for facial expressions analysis using dynamic and static information in video sequences. First, based on incremental formulation, discriminative deformable face alignment method is adapted to locate facial points to correct in-plane head rotation and break up facial region from background. Then, spatial-temporal motion local binary pattern (LBP feature is extracted and integrated with Gabor multiorientation fusion histogram to give descriptors, which reflect static and dynamic texture information of facial expressions. Finally, a one-versus-one strategy based multiclass support vector machine (SVM classifier is applied to classify facial expressions. Experiments on Cohn-Kanade (CK + facial expression dataset illustrate that integrated framework outperforms methods using single descriptors. Compared with other state-of-the-art methods on CK+, MMI, and Oulu-CASIA VIS datasets, our proposed framework performs better.

  3. Probabilistic model based error correction in a set of various mutant sequences analyzed by next-generation sequencing.

    Science.gov (United States)

    Aita, Takuyo; Ichihashi, Norikazu; Yomo, Tetsuya

    2013-12-01

    To analyze the evolutionary dynamics of a mutant population in an evolutionary experiment, it is necessary to sequence a vast number of mutants by high-throughput (next-generation) sequencing technologies, which enable rapid and parallel analysis of multikilobase sequences. However, the observed sequences include many errors of base call. Therefore, if next-generation sequencing is applied to analysis of a heterogeneous population of various mutant sequences, it is necessary to discriminate between true bases as point mutations and errors of base call in the observed sequences, and to subject the sequences to error-correction processes. To address this issue, we have developed a novel method of error correction based on the Potts model and a maximum a posteriori probability (MAP) estimate of its parameters corresponding to the "true sequences". Our method of error correction utilizes (1) the "quality scores" which are assigned to individual bases in the observed sequences and (2) the neighborhood relationship among the observed sequences mapped in sequence space. The computer experiments of error correction of artificially generated sequences supported the effectiveness of our method, showing that 50-90% of errors were removed. Interestingly, this method is analogous to a probabilistic model based method of image restoration developed in the field of information engineering.

  4. Visual programming for next-generation sequencing data analytics.

    Science.gov (United States)

    Milicchio, Franco; Rose, Rebecca; Bian, Jiang; Min, Jae; Prosperi, Mattia

    2016-01-01

    High-throughput or next-generation sequencing (NGS) technologies have become an established and affordable experimental framework in biological and medical sciences for all basic and translational research. Processing and analyzing NGS data is challenging. NGS data are big, heterogeneous, sparse, and error prone. Although a plethora of tools for NGS data analysis has emerged in the past decade, (i) software development is still lagging behind data generation capabilities, and (ii) there is a 'cultural' gap between the end user and the developer. Generic software template libraries specifically developed for NGS can help in dealing with the former problem, whilst coupling template libraries with visual programming may help with the latter. Here we scrutinize the state-of-the-art low-level software libraries implemented specifically for NGS and graphical tools for NGS analytics. An ideal developing environment for NGS should be modular (with a native library interface), scalable in computational methods (i.e. serial, multithread, distributed), transparent (platform-independent), interoperable (with external software interface), and usable (via an intuitive graphical user interface). These characteristics should facilitate both the run of standardized NGS pipelines and the development of new workflows based on technological advancements or users' needs. We discuss in detail the potential of a computational framework blending generic template programming and visual programming that addresses all of the current limitations. In the long term, a proper, well-developed (although not necessarily unique) software framework will bridge the current gap between data generation and hypothesis testing. This will eventually facilitate the development of novel diagnostic tools embedded in routine healthcare.

  5. Influence of the Choice of Core-Envelope Transition Point on the Binary Merger of Two Main-sequence Components

    Institute of Scientific and Technical Information of China (English)

    Xue-Fei Chen; Zhan-Wen Han

    2005-01-01

    We have studied the influence of different choices of core-envelope transition point on the final merger of contact binaries with two main-sequence components. A binary of 1.00 + 0.90 M⊙ with an initial orbital period of 0.35d is examined. The mass fraction of the primary mixed with the matter of the secondary,qmix, determined by the chosen core-envelope transition point, ranges from 0.04 to 1.00 in our analysis. If as qmix < 0.8, none of the helium-rich matter in the center of the primary is mixed into the envelope, and there is little distinction in the evolutionary tracks of the mergers. The timescales of the mergers remaining on the main sequence, tBS, are very similar (~ 6.2 × 108yr) if qmix < 0.71, since no hydrogen-rich matter of the secondary is mixed into the core of the mergers;for qmix > 0.71, the larger qmix is, the greater the mixing, hence the longer the blue straggler lifetime, tBS, and also the greater the luminosity. For qmix = 1.00,tBS ~ 8.5 × 10/ yr. Estimation by ( )r - ( )a = 0.0 shows that the point at which tBS begins to increase is about qmix = 0.68. In comparison with the homogeneously mixed models, the merger with a helium profile similar to that of the primary is less luminous and has a shorter tBS.

  6. A program for generating randomized simple and context-sensitive sequences.

    Science.gov (United States)

    Remillard, Gilbert

    2008-05-01

    This article introduces Sequence Generation 2008 (SeqGen2008), a Windows-based sequence generator. SeqGen2008 can generate simple sequences satisfying user-defined event probabilities or frequencies. The program can also generate context-sensitive sequences satisfying user-defined transition matrices that specify the probabilities or frequencies with which distinct events are to follow specific contexts. An analysis of the properties and behavior of the algorithms employed by SeqGen2008 reveals that the algorithms are unbiased in their generation of sequences.

  7. Authentication of Herbal Supplements Using Next-Generation Sequencing.

    Directory of Open Access Journals (Sweden)

    Natalia V Ivanova

    Full Text Available DNA-based testing has been gaining acceptance as a tool for authentication of a wide range of food products; however, its applicability for testing of herbal supplements remains contentious.We utilized Sanger and Next-Generation Sequencing (NGS for taxonomic authentication of fifteen herbal supplements representing three different producers from five medicinal plants: Echinacea purpurea, Valeriana officinalis, Ginkgo biloba, Hypericum perforatum and Trigonella foenum-graecum. Experimental design included three modifications of DNA extraction, two lysate dilutions, Internal Amplification Control, and multiple negative controls to exclude background contamination. Ginkgo supplements were also analyzed using HPLC-MS for the presence of active medicinal components.All supplements yielded DNA from multiple species, rendering Sanger sequencing results for rbcL and ITS2 regions either uninterpretable or non-reproducible between the experimental replicates. Overall, DNA from the manufacturer-listed medicinal plants was successfully detected in seven out of eight dry herb form supplements; however, low or poor DNA recovery due to degradation was observed in most plant extracts (none detected by Sanger; three out of seven-by NGS. NGS also revealed a diverse community of fungi, known to be associated with live plant material and/or the fermentation process used in the production of plant extracts. HPLC-MS testing demonstrated that Ginkgo supplements with degraded DNA contained ten key medicinal components.Quality control of herbal supplements should utilize a synergetic approach targeting both DNA and bioactive components, especially for standardized extracts with degraded DNA. The NGS workflow developed in this study enables reliable detection of plant and fungal DNA and can be utilized by manufacturers for quality assurance of raw plant materials, contamination control during the production process, and the final product. Interpretation of results should

  8. Autocorrelation of Sequences Generated by Single Cycle T-Functions

    Institute of Scientific and Technical Information of China (English)

    Wang Yan; Hu Yupu; Li Shunbo; Yang Yang

    2011-01-01

    Cryptographic properties of the single cycle T-function's output sequences are investigated.Bounds of autocorrelation functions of the kth coordinate sequence and bounds of state output sequence are calculated respectively.The Maximum Sidelobe Ratio (MSR) of the kth coordinate sequence and the MSR of state output sequence are given respectively.The bounds of autocorrelation functions show that the values of autocorrelation functions are large when shifts are small.Comparisons of the autocorrelations between the state output sequence and coordinate output sequence are illustrated.The autocorrelation properties demonstrate that T-functions have cryptographic weaknesses and the illustration result shows coordinate output sequences have better autocorrelation than that of state output sequences.

  9. MML 53: a new low-mass, pre-main sequence eclipsing binary in the Upper Centarus-Lupus Region discovered by SuperWASP

    CERN Document Server

    Hebb, L; Aigrain, S; Collier-Cameron, A; Hodgkin, S T; Irwin, J M; Maxted, P F L; Pollacco, D; Street, R A; Wilson, D M; Stassun, K G

    2010-01-01

    We announce the discovery of a new low-mass, pre-main sequence eclipsing binary, MML 53. Previous observations of MML 53 found it to be a pre-main sequence spectroscopic multiple associated with the 15-22 Myr Upper Centaurus Lupus cluster. We identify the object as an eclipsing binary for the first time through the analysis of multiple seasons of time series photometry from the SuperWASP transiting planet survey. Re-analysis of a single archive spectrum shows MML 53 to be a spatially unresolved triple system of young stars which all exhibit significant lithium absorption. Two of the components comprise an eclipsing binary with period, P = 2.097891(6) +- 0.000005 and mass ratio, q~0.8. Here, we present the analysis of the discovery data.

  10. DNA qualification workflow for next generation sequencing of histopathological samples.

    Directory of Open Access Journals (Sweden)

    Michele Simbolo

    Full Text Available Histopathological samples are a treasure-trove of DNA for clinical research. However, the quality of DNA can vary depending on the source or extraction method applied. Thus a standardized and cost-effective workflow for the qualification of DNA preparations is essential to guarantee interlaboratory reproducible results. The qualification process consists of the quantification of double strand DNA (dsDNA and the assessment of its suitability for downstream applications, such as high-throughput next-generation sequencing. We tested the two most frequently used instrumentations to define their role in this process: NanoDrop, based on UV spectroscopy, and Qubit 2.0, which uses fluorochromes specifically binding dsDNA. Quantitative PCR (qPCR was used as the reference technique as it simultaneously assesses DNA concentration and suitability for PCR amplification. We used 17 genomic DNAs from 6 fresh-frozen (FF tissues, 6 formalin-fixed paraffin-embedded (FFPE tissues, 3 cell lines, and 2 commercial preparations. Intra- and inter-operator variability was negligible, and intra-methodology variability was minimal, while consistent inter-methodology divergences were observed. In fact, NanoDrop measured DNA concentrations higher than Qubit and its consistency with dsDNA quantification by qPCR was limited to high molecular weight DNA from FF samples and cell lines, where total DNA and dsDNA quantity virtually coincide. In partially degraded DNA from FFPE samples, only Qubit proved highly reproducible and consistent with qPCR measurements. Multiplex PCR amplifying 191 regions of 46 cancer-related genes was designated the downstream application, using 40 ng dsDNA from FFPE samples calculated by Qubit. All but one sample produced amplicon libraries suitable for next-generation sequencing. NanoDrop UV-spectrum verified contamination of the unsuccessful sample. In conclusion, as qPCR has high costs and is labor intensive, an alternative effective standard

  11. DNA qualification workflow for next generation sequencing of histopathological samples.

    Science.gov (United States)

    Simbolo, Michele; Gottardi, Marisa; Corbo, Vincenzo; Fassan, Matteo; Mafficini, Andrea; Malpeli, Giorgio; Lawlor, Rita T; Scarpa, Aldo

    2013-01-01

    Histopathological samples are a treasure-trove of DNA for clinical research. However, the quality of DNA can vary depending on the source or extraction method applied. Thus a standardized and cost-effective workflow for the qualification of DNA preparations is essential to guarantee interlaboratory reproducible results. The qualification process consists of the quantification of double strand DNA (dsDNA) and the assessment of its suitability for downstream applications, such as high-throughput next-generation sequencing. We tested the two most frequently used instrumentations to define their role in this process: NanoDrop, based on UV spectroscopy, and Qubit 2.0, which uses fluorochromes specifically binding dsDNA. Quantitative PCR (qPCR) was used as the reference technique as it simultaneously assesses DNA concentration and suitability for PCR amplification. We used 17 genomic DNAs from 6 fresh-frozen (FF) tissues, 6 formalin-fixed paraffin-embedded (FFPE) tissues, 3 cell lines, and 2 commercial preparations. Intra- and inter-operator variability was negligible, and intra-methodology variability was minimal, while consistent inter-methodology divergences were observed. In fact, NanoDrop measured DNA concentrations higher than Qubit and its consistency with dsDNA quantification by qPCR was limited to high molecular weight DNA from FF samples and cell lines, where total DNA and dsDNA quantity virtually coincide. In partially degraded DNA from FFPE samples, only Qubit proved highly reproducible and consistent with qPCR measurements. Multiplex PCR amplifying 191 regions of 46 cancer-related genes was designated the downstream application, using 40 ng dsDNA from FFPE samples calculated by Qubit. All but one sample produced amplicon libraries suitable for next-generation sequencing. NanoDrop UV-spectrum verified contamination of the unsuccessful sample. In conclusion, as qPCR has high costs and is labor intensive, an alternative effective standard workflow for

  12. Automated Generation of Phase Diagrams for Binary Systems with Azeotropic Behavior

    DEFF Research Database (Denmark)

    Cismondi, Martin; Michelsen, Michael Locht; Zabaloy, Marcelo S.

    2008-01-01

    In this work, we propose a computational strategy and methods for the automated calculation of complete loci of homogeneous azeotropy of binary mixtures and the related Pxy and Txy diagrams for models of the equation-of-state (EOS) type. The strategy consists of first finding the system's azeotro...

  13. DISCUSSION ON AVERAGE GENERATION OF TWODIMENSIONAL DATA SEQUENCE IN GREY SYSTEM THEORY

    Institute of Scientific and Technical Information of China (English)

    PINGXue-liang; ZHOURu-rong; LIUSheng-lan

    2004-01-01

    An unequal time interval sequence or a sequence with blanks is usually completed with average generation in grey system theory. This paper discovers that there exists obvious errors when using average generation to generate internal points of non-consecutive neighbours. The average generation and the preference generation of the sequence are discussed, the concave and convex properties show the status of local sequence and propose a new idea for using the status to build up the criteria of choosing generation coefficient. Compared with the general average method of the one-dimensional data sequence, the two-dimensional data sequence is defined and its average generation is discussed, and the coefficient decision method for the preference generation is presented.

  14. The rolling circle amplification and next generation sequencing ...

    African Journals Online (AJOL)

    Titus

    2016-09-14

    Sep 14, 2016 ... sequencing approaches reveal genome wide diversity of Kenyan cassava mosaic ..... Kenya for providing the funds for this project. Our special ... (2008). Accurate whole human genome sequencing using reversible terminator ...

  15. Peptide Synthesis on a Next-Generation DNA Sequencing Platform.

    Science.gov (United States)

    Svensen, Nina; Peersen, Olve B; Jaffrey, Samie R

    2016-09-01

    Methods for displaying large numbers of peptides on solid surfaces are essential for high-throughput characterization of peptide function and binding properties. Here we describe a method for converting the >10(7) flow cell-bound clusters of identical DNA strands generated by the Illumina DNA sequencing technology into clusters of complementary RNA, and subsequently peptide clusters. We modified the flow-cell-bound primers with ribonucleotides thus enabling them to be used by poliovirus polymerase 3D(pol) . The primers hybridize to the clustered DNA thus leading to RNA clusters. The RNAs fold into functional protein- or small molecule-binding aptamers. We used the mRNA-display approach to synthesize flow-cell-tethered peptides from these RNA clusters. The peptides showed selective binding to cognate antibodies. The methods described here provide an approach for using DNA clusters to template peptide synthesis on an Illumina flow cell, thus providing new opportunities for massively parallel peptide-based assays.

  16. Accelerating next generation sequencing data analysis with system level optimizations.

    Science.gov (United States)

    Kathiresan, Nagarajan; Temanni, Ramzi; Almabrazi, Hakeem; Syed, Najeeb; Jithesh, Puthen V; Al-Ali, Rashid

    2017-08-22

    Next generation sequencing (NGS) data analysis is highly compute intensive. In-memory computing, vectorization, bulk data transfer, CPU frequency scaling are some of the hardware features in the modern computing architectures. To get the best execution time and utilize these hardware features, it is necessary to tune the system level parameters before running the application. We studied the GATK-HaplotypeCaller which is part of common NGS workflows, that consume more than 43% of the total execution time. Multiple GATK 3.x versions were benchmarked and the execution time of HaplotypeCaller was optimized by various system level parameters which included: (i) tuning the parallel garbage collection and kernel shared memory to simulate in-memory computing, (ii) architecture-specific tuning in the PairHMM library for vectorization, (iii) including Java 1.8 features through GATK source code compilation and building a runtime environment for parallel sorting and bulk data transfer (iv) the default 'on-demand' mode of CPU frequency is over-clocked by using 'performance-mode' to accelerate the Java multi-threads. As a result, the HaplotypeCaller execution time was reduced by 82.66% in GATK 3.3 and 42.61% in GATK 3.7. Overall, the execution time of NGS pipeline was reduced to 70.60% and 34.14% for GATK 3.3 and GATK 3.7 respectively.

  17. Deep homology in the age of next-generation sequencing.

    Science.gov (United States)

    Tschopp, Patrick; Tabin, Clifford J

    2017-02-05

    The principle of homology is central to conceptualizing the comparative aspects of morphological evolution. The distinctions between homologous or non-homologous structures have become blurred, however, as modern evolutionary developmental biology (evo-devo) has shown that novel features often result from modification of pre-existing developmental modules, rather than arising completely de novo. With this realization in mind, the term 'deep homology' was coined, in recognition of the remarkably conserved gene expression during the development of certain animal structures that would not be considered homologous by previous strict definitions. At its core, it can help to formulate an understanding of deeper layers of ontogenetic conservation for anatomical features that lack any clear phylogenetic continuity. Here, we review deep homology and related concepts in the context of a gene expression-based homology discussion. We then focus on how these conceptual frameworks have profited from the recent rise of high-throughput next-generation sequencing. These techniques have greatly expanded the range of organisms amenable to such studies. Moreover, they helped to elevate the traditional gene-by-gene comparison to a transcriptome-wide level. We will end with an outlook on the next challenges in the field and how technological advances might provide exciting new strategies to tackle these questions.This article is part of the themed issue 'Evo-devo in the genomics era, and the origins of morphological diversity'. © 2016 The Author(s).

  18. Next-generation sequence assembly: four stages of data processing and computational challenges.

    Directory of Open Access Journals (Sweden)

    Sara El-Metwally

    Full Text Available Decoding DNA symbols using next-generation sequencers was a major breakthrough in genomic research. Despite the many advantages of next-generation sequencers, e.g., the high-throughput sequencing rate and relatively low cost of sequencing, the assembly of the reads produced by these sequencers still remains a major challenge. In this review, we address the basic framework of next-generation genome sequence assemblers, which comprises four basic stages: preprocessing filtering, a graph construction process, a graph simplification process, and postprocessing filtering. Here we discuss them as a framework of four stages for data analysis and processing and survey variety of techniques, algorithms, and software tools used during each stage. We also discuss the challenges that face current assemblers in the next-generation environment to determine the current state-of-the-art. We recommend a layered architecture approach for constructing a general assembler that can handle the sequences generated by different sequencing platforms.

  19. Next-generation sequence assembly: four stages of data processing and computational challenges.

    Science.gov (United States)

    El-Metwally, Sara; Hamza, Taher; Zakaria, Magdi; Helmy, Mohamed

    2013-01-01

    Decoding DNA symbols using next-generation sequencers was a major breakthrough in genomic research. Despite the many advantages of next-generation sequencers, e.g., the high-throughput sequencing rate and relatively low cost of sequencing, the assembly of the reads produced by these sequencers still remains a major challenge. In this review, we address the basic framework of next-generation genome sequence assemblers, which comprises four basic stages: preprocessing filtering, a graph construction process, a graph simplification process, and postprocessing filtering. Here we discuss them as a framework of four stages for data analysis and processing and survey variety of techniques, algorithms, and software tools used during each stage. We also discuss the challenges that face current assemblers in the next-generation environment to determine the current state-of-the-art. We recommend a layered architecture approach for constructing a general assembler that can handle the sequences generated by different sequencing platforms.

  20. 白矮-主序双星的搜寻及研究进展%Research Progress on Searching for White Dwarf-Main Sequence Binaries

    Institute of Scientific and Technical Information of China (English)

    任娟娟; 罗阿理; 赵永恒

    2014-01-01

    White dwarf-main sequence binaries (WDMS) are the most common compact binary ob jects in the Galaxy, each of which consists of a white dwarf and a main sequence star and is evolved from main sequence binary. About 25 percent of the WDMS binaries are close WDMS binaries that evolved through a common envelope phase, and are commonly referred to as post-common-envelope binaries (PCEBs). The remaining 75 percent are wide WDMS binaries that did not evolve through a common envelope phase, with the orbital separation roughly the same as the orbital separation of the initial main sequence binary. Generally, the two components can be seen clearly from the WDMS binary spectra optically. Thanks to the large spectroscopic survey like SDSS and LAMOST, the number of WDMS binaries has been increased dramatically recently. A large number of wide WDMS binaries and PCEBs have been identified by the follow-up observations of these WDMS bina-ries. Currently, more than 2000 WDMS binaries have been discovered spectroscopically and about 200 PCEBs have been confirmed. Upon the large sample of SDSS WDMS binaries and PCEBs identified, many important researches have been carried on, such as the com-mon envelope theory, the origin of low mass white dwarf, mass-radius relations of both white dwarfs and low mass main sequence stars, and the pairing properties of main sequence stars. However, as the SDSS WDMS binaries sample has serious selection effects, which is strongly biased against binary systems containing cool white dwarf and/or early type companions, we still need to search more WDMS binaries to enlarge the sample. The LAMOST sky survey began its five years regular survey from September 2012, which will observe a large number of targets in the Milky Way. From the recent data release (DR1) of LAMOST, more than 100 WDMS binaries have been found. With the ongoing SDSS and LAMOST survey, more WDMS binaries are hoped to be identified and extend the existing WDMS binary sample. In this paper

  1. Sequence Analysis of Trimer Isomers Formed by Montmorillonite Catalysis in the Reaction of Binary Monomer Mixtures

    Science.gov (United States)

    Ertem, Gözen; Hazen, Robert M.; Dworkin, Jason P.

    2007-10-01

    Oligonucleotides are structurally similar to short RNA strands. Therefore, their formation via non-enzymatic reactions is highly relevant to Gilbert's RNA world scenario (1986) and the origin of life. In laboratory synthesis of oligonucleotides from monomers, it is necessary to remove the water molecules from the reaction medium to shift the equilibrium in favor of oligonucleotide formation, which would have been impossible for reactions that took place in dilute solutions on the early Earth. Model studies designed to address this problem demonstrate that montmorillonite, a phyllosilicate common on Earth and identified on Mars, efficiently catalyzes phosphodiester-bond formation between activated mononucleotides in dilute solutions and produces RNA-like oligomers. The purpose of this study was to examine the sequences and regiospecificity of trimer isomers formed in the reaction of 5'-phosphorimidazolides of adenosine and uridine. Results demonstrated that regiospecificity and sequence specificity observed in the dimer fractions are conserved in their elongation products. With regard to regiospecificity, 61% of the linkages were found to be RNA-like 3',5'-phosphodiester bonds. With regard to sequence specificity, we found that 88% of the linear trimers were hetero-isomers with 61% A-monomer and 39% U-monomer incorporation. These results lend support to Bernal's hypothesis that minerals may have played a significant role in the chemical processes that led to the origin of life by catalyzing the formation of phosphodiester bonds in RNA-like oligomers.

  2. Next-generation DNA barcoding: using next-generation sequencing to enhance and accelerate DNA barcode capture from single specimens.

    Science.gov (United States)

    Shokralla, Shadi; Gibson, Joel F; Nikbakht, Hamid; Janzen, Daniel H; Hallwachs, Winnie; Hajibabaei, Mehrdad

    2014-09-01

    DNA barcoding is an efficient method to identify specimens and to detect undescribed/cryptic species. Sanger sequencing of individual specimens is the standard approach in generating large-scale DNA barcode libraries and identifying unknowns. However, the Sanger sequencing technology is, in some respects, inferior to next-generation sequencers, which are capable of producing millions of sequence reads simultaneously. Additionally, direct Sanger sequencing of DNA barcode amplicons, as practiced in most DNA barcoding procedures, is hampered by the need for relatively high-target amplicon yield, coamplification of nuclear mitochondrial pseudogenes, confusion with sequences from intracellular endosymbiotic bacteria (e.g. Wolbachia) and instances of intraindividual variability (i.e. heteroplasmy). Any of these situations can lead to failed Sanger sequencing attempts or ambiguity of the generated DNA barcodes. Here, we demonstrate the potential application of next-generation sequencing platforms for parallel acquisition of DNA barcode sequences from hundreds of specimens simultaneously. To facilitate retrieval of sequences obtained from individual specimens, we tag individual specimens during PCR amplification using unique 10-mer oligonucleotides attached to DNA barcoding PCR primers. We employ 454 pyrosequencing to recover full-length DNA barcodes of 190 specimens using 12.5% capacity of a 454 sequencing run (i.e. two lanes of a 16 lane run). We obtained an average of 143 sequence reads for each individual specimen. The sequences produced are full-length DNA barcodes for all but one of the included specimens. In a subset of samples, we also detected Wolbachia, nontarget species, and heteroplasmic sequences. Next-generation sequencing is of great value because of its protocol simplicity, greatly reduced cost per barcode read, faster throughout and added information content.

  3. Using Layer Recurrent Neural Network to Generate Pseudo Random Number Sequences

    Directory of Open Access Journals (Sweden)

    Veena Desai

    2012-03-01

    Full Text Available Pseudo Random Numbers (PRNs are required for many cryptographic applications. This paper proposes a new method for generating PRNs using Layer Recurrent Neural Network (LRNN. The proposed technique generates PRNs from the weight matrix obtained from the layer weights of the LRNN. The LRNN random number generator (RNG uses a short keyword as a seed and generates a long sequence as a pseudo PRN sequence. The number of bits generated in the PRN sequence depends on the number of neurons in the input layer of the LRNN. The generated PRN sequence changes, with a change in the training function of the LRNN .The sequences generated are a function of the keyword, initial state of network and the training function. In our implementation the PRN sequences have been generated using 3 training functions: 1Scaled Gradient Descent 2Levenberg-Marquartz (TRAINLM and 3 TRAINBGF. The generated sequences are tested for randomness using ENT and NIST test suites. The ENT test can be applied for sequences of small size. NIST has 16 tests to test random numbers. The LRNN generated PRNs pass in 11 tests, show no observations for 4 tests, and fail in 1 test when subjected to NIST .This paper presents the test results for random number sequence ranging from 25 bits to 1000 bits, generated using LRNN.

  4. Next generation sequencing (NGS)technologies and applications

    Energy Technology Data Exchange (ETDEWEB)

    Vuyisich, Momchilo [Los Alamos National Laboratory

    2012-09-11

    NGS technology overview: (1) NGS library preparation - Nucleic acids extraction, Sample quality control, RNA conversion to cDNA, Addition of sequencing adapters, Quality control of library; (2) Sequencing - Clonal amplification of library fragments, (except PacBio), Sequencing by synthesis, Data output (reads and quality); and (3) Data analysis - Read mapping, Genome assembly, Gene expression, Operon structure, sRNA discovery, and Epigenetic analyses.

  5. Dynamo generated magnetic configurations in accretion discs and the nature of quasi-periodic oscillations in accreting binary systems

    CERN Document Server

    Moss, David; Suleimanov, Valery

    2016-01-01

    Magnetic fields are important for accretion disc structure. Magnetic fields in a disc system may be transported with the accreted matter. They can be associated with either the central body and/or jet, and be fossil or dynamo excited in situ. We consider dynamo excitation of magnetic fields in accretion discs of accreting binary systems in an attempt to clarify possible configurations of dynamo generated magnetic fields. We first model the entire disc with realistic radial extent and thickness using an alpha-quenching non-linearity. We then study the simultaneous effect of feedback from the Lorentz force from the dynamo-generated field. We perform numerical simulations in the framework of a relatively simple mean-field model which allows the generation of global magnetic configurations. We explore a range of possibilities for the dynamo number, and find quadrupolar-type solutions with irregular temporal oscillations that might be compared to observed rapid luminosity fluctuations. The dipolar symmetry models ...

  6. Development of hydrothermal power generation plant. Development of binary cycle power generation plant (development of 10 MW-class plant); 1995 nendo nessui riyo hatsuden plant nado kaihatsu binary cycle hatsuden plant no kaihatsu. 10MW kyu plant no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-12-01

    A 10 MW-class binary cycle power generation plant has been developed using a down hole pump (DHP) which exchanges the hydrothermal energy with secondary medium in the heat exchanger. For constructing the plant at Kuju-machi, Oita Prefecture, site preparation works, foundation of cooling tower, reconstruction of roads, and survey on environmental influences were conducted. To investigate installation and removal methods of DHP, a geothermal water pump-up system, current status of the binary cycle power generating system in the USA was surveyed. In this survey, a trailer mounting handling machine was inspected. Based on the survey results, a simple assembled, easy-installation type handling equipment was designed. In addition, the replacement work for motor connector joint of DHP and the strength of coil end were improved. Construction and method allowing reuse of the motor cable were considered by improving the cable and cable end portion. The air tight soundness of incoloy corrugate sheath was confirmed. Finally, a reproduction system for waste oil of DHP bearing oil was investigated. 106 figs., 52 tabs.

  7. Historical Perspective, Development and Applications of Next-Generation Sequencing in Plant Virology

    OpenAIRE

    Marina Barba; Henryk Czosnek; Ahmed Hadidi

    2014-01-01

    Next-generation high throughput sequencing technologies became available at the onset of the 21st century. They provide a highly efficient, rapid, and low cost DNA sequencing platform beyond the reach of the standard and traditional DNA sequencing technologies developed in the late 1970s. They are continually improved to become faster, more efficient and cheaper. They have been used in many fields of biology since 2004. In 2009, next-generation sequencing (NGS) technologies began to be appli...

  8. Applications of Next-Generation Sequencing Technologies to Diagnostic Virology

    Directory of Open Access Journals (Sweden)

    Giorgio Palù

    2011-11-01

    Full Text Available Novel DNA sequencing techniques, referred to as “next-generation” sequencing (NGS, provide high speed and throughput that can produce an enormous volume of sequences with many possible applications in research and diagnostic settings. In this article, we provide an overview of the many applications of NGS in diagnostic virology. NGS techniques have been used for high-throughput whole viral genome sequencing, such as sequencing of new influenza viruses, for detection of viral genome variability and evolution within the host, such as investigation of human immunodeficiency virus and human hepatitis C virus quasispecies, and monitoring of low-abundance antiviral drug-resistance mutations. NGS techniques have been applied to metagenomics-based strategies for the detection of unexpected disease-associated viruses and for the discovery of novel human viruses, including cancer-related viruses. Finally, the human virome in healthy and disease conditions has been described by NGS-based metagenomics.

  9. CAPRG: sequence assembling pipeline for next generation sequencing of non-model organisms.

    Directory of Open Access Journals (Sweden)

    Arun Rawat

    Full Text Available Our goal is to introduce and describe the utility of a new pipeline "Contigs Assembly Pipeline using Reference Genome" (CAPRG, which has been developed to assemble "long sequence reads" for non-model organisms by leveraging a reference genome of a closely related phylogenetic relative. To facilitate this effort, we utilized two avian transcriptomic datasets generated using ROCHE/454 technology as test cases for CAPRG assembly. We compared the results of CAPRG assembly using a reference genome with the results of existing methods that utilize de novo strategies such as VELVET, PAVE, and MIRA by employing parameter space comparisons (intra-assembling comparison. CAPRG performed as well or better than the existing assembly methods based on various benchmarks for "gene-hunting." Further, CAPRG completed the assemblies in a fraction of the time required by the existing assembly algorithms. Additional advantages of CAPRG included reduced contig inflation resulting in lower computational resources for annotation, and functional identification for contigs that may be categorized as "unknowns" by de novo methods. In addition to providing evaluation of CAPRG performance, we observed that the different assembly (inter-assembly results could be integrated to enhance the putative gene coverage for any transcriptomics study.

  10. CAPRG: sequence assembling pipeline for next generation sequencing of non-model organisms.

    Science.gov (United States)

    Rawat, Arun; Elasri, Mohamed O; Gust, Kurt A; George, Glover; Pham, Don; Scanlan, Leona D; Vulpe, Chris; Perkins, Edward J

    2012-01-01

    Our goal is to introduce and describe the utility of a new pipeline "Contigs Assembly Pipeline using Reference Genome" (CAPRG), which has been developed to assemble "long sequence reads" for non-model organisms by leveraging a reference genome of a closely related phylogenetic relative. To facilitate this effort, we utilized two avian transcriptomic datasets generated using ROCHE/454 technology as test cases for CAPRG assembly. We compared the results of CAPRG assembly using a reference genome with the results of existing methods that utilize de novo strategies such as VELVET, PAVE, and MIRA by employing parameter space comparisons (intra-assembling comparison). CAPRG performed as well or better than the existing assembly methods based on various benchmarks for "gene-hunting." Further, CAPRG completed the assemblies in a fraction of the time required by the existing assembly algorithms. Additional advantages of CAPRG included reduced contig inflation resulting in lower computational resources for annotation, and functional identification for contigs that may be categorized as "unknowns" by de novo methods. In addition to providing evaluation of CAPRG performance, we observed that the different assembly (inter-assembly) results could be integrated to enhance the putative gene coverage for any transcriptomics study.

  11. Targeted next-generation sequencing can replace Sanger sequencing in clinical diagnostics

    NARCIS (Netherlands)

    Sikkema-Raddatz, B.; Johansson, L.F.; de Boer, E.N.; Almomani, R.; Boven, L.G.; van den Berg, M.P.; van Spaendonck-Zwarts, K.Y.; van Tintelen, J.P.; Sijmons, R.H.; Jongbloed, J.D.H.; Sinke, R.J.

    2013-01-01

    Mutation detection through exome sequencing allows simultaneous analysis of all coding sequences of genes. However, it cannot yet replace Sanger sequencing (SS) in diagnostics because of incomplete representation and coverage of exons leading to missing clinically relevant mutations. Targeted next-g

  12. Recurrent Network Models of Sequence Generation and Memory.

    Science.gov (United States)

    Rajan, Kanaka; Harvey, Christopher D; Tank, David W

    2016-04-01

    Sequential activation of neurons is a common feature of network activity during a variety of behaviors, including working memory and decision making. Previous network models for sequences and memory emphasized specialized architectures in which a principled mechanism is pre-wired into their connectivity. Here we demonstrate that, starting from random connectivity and modifying a small fraction of connections, a largely disordered recurrent network can produce sequences and implement working memory efficiently. We use this process, called Partial In-Network Training (PINning), to model and match cellular resolution imaging data from the posterior parietal cortex during a virtual memory-guided two-alternative forced-choice task. Analysis of the connectivity reveals that sequences propagate by the cooperation between recurrent synaptic interactions and external inputs, rather than through feedforward or asymmetric connections. Together our results suggest that neural sequences may emerge through learning from largely unstructured network architectures.

  13. Parameter estimation for compact binary coalescence signals with the first generation gravitational-wave detector network

    Science.gov (United States)

    Aasi, J.; Abadie, J.; Abbott, B. P.; Abbott, R.; Abbott, T. D.; Abernathy, M.; Accadia, T.; Acernese, F.; Adams, C.; Adams, T.; Addesso, P.; Adhikari, R.; Affeldt, C.; Agathos, M.; Agatsuma, K.; Ajith, P.; Allen, B.; Allocca, A.; Amador Ceron, E.; Amariutei, D.; Anderson, S. B.; Anderson, W. G.; Arai, K.; Araya, M. C.; Ast, S.; Aston, S. M.; Astone, P.; Atkinson, D.; Aufmuth, P.; Aulbert, C.; Aylott, B. E.; Babak, S.; Baker, P.; Ballardin, G.; Ballmer, S.; Bao, Y.; Barayoga, J. C. B.; Barker, D.; Barone, F.; Barr, B.; Barsotti, L.; Barsuglia, M.; Barton, M. A.; Bartos, I.; Bassiri, R.; Bastarrika, M.; Basti, A.; Batch, J.; Bauchrowitz, J.; Bauer, Th. S.; Bebronne, M.; Beck, D.; Behnke, B.; Bejger, M.; Beker, M. G.; Bell, A. S.; Bell, C.; Belopolski, I.; Benacquista, M.; Berliner, J. M.; Bertolini, A.; Betzwieser, J.; Beveridge, N.; Beyersdorf, P. T.; Bhadbade, T.; Bilenko, I. A.; Billingsley, G.; Birch, J.; Biswas, R.; Bitossi, M.; Bizouard, M. A.; Black, E.; Blackburn, J. K.; Blackburn, L.; Blair, D.; Bland, B.; Blom, M.; Bock, O.; Bodiya, T. P.; Bogan, C.; Bond, C.; Bondarescu, R.; Bondu, F.; Bonelli, L.; Bonnand, R.; Bork, R.; Born, M.; Boschi, V.; Bose, S.; Bosi, L.; Bouhou, B.; Braccini, S.; Bradaschia, C.; Brady, P. R.; Braginsky, V. B.; Branchesi, M.; Brau, J. E.; Breyer, J.; Briant, T.; Bridges, D. O.; Brillet, A.; Brinkmann, M.; Brisson, V.; Britzger, M.; Brooks, A. F.; Brown, D. A.; Bulik, T.; Bulten, H. J.; Buonanno, A.; Burguet–Castell, J.; Buskulic, D.; Buy, C.; Byer, R. L.; Cadonati, L.; Cagnoli, G.; Calloni, E.; Camp, J. B.; Campsie, P.; Cannon, K.; Canuel, B.; Cao, J.; Capano, C. D.; Carbognani, F.; Carbone, L.; Caride, S.; Caudill, S.; Cavaglià, M.; Cavalier, F.; Cavalieri, R.; Cella, G.; Cepeda, C.; Cesarini, E.; Chalermsongsak, T.; Charlton, P.; Chassande-Mottin, E.; Chen, W.; Chen, X.; Chen, Y.; Chincarini, A.; Chiummo, A.; Cho, H. S.; Chow, J.; Christensen, N.; Chua, S. S. Y.; Chung, C. T. Y.; Chung, S.; Ciani, G.; Clara, F.; Clark, D. E.; Clark, J. A.; Clayton, J. H.; Cleva, F.; Coccia, E.; Cohadon, P.-F.; Colacino, C. N.; Colla, A.; Colombini, M.; Conte, A.; Conte, R.; Cook, D.; Corbitt, T. R.; Cordier, M.; Cornish, N.; Corsi, A.; Costa, C. A.; Coughlin, M.; Coulon, J.-P.; Couvares, P.; Coward, D. M.; Cowart, M.; Coyne, D. C.; Creighton, J. D. E.; Creighton, T. D.; Cruise, A. M.; Cumming, A.; Cunningham, L.; Cuoco, E.; Cutler, R. M.; Dahl, K.; Damjanic, M.; Danilishin, S. L.; D'Antonio, S.; Danzmann, K.; Dattilo, V.; Daudert, B.; Daveloza, H.; Davier, M.; Daw, E. J.; Dayanga, T.; De Rosa, R.; DeBra, D.; Debreczeni, G.; Degallaix, J.; Del Pozzo, W.; Dent, T.; Dergachev, V.; DeRosa, R.; Dhurandhar, S.; Di Fiore, L.; Di Lieto, A.; Di Palma, I.; Di Paolo Emilio, M.; Di Virgilio, A.; Díaz, M.; Dietz, A.; Donovan, F.; Dooley, K. L.; Doravari, S.; Dorsher, S.; Drago, M.; Drever, R. W. P.; Driggers, J. C.; Du, Z.; Dumas, J.-C.; Dwyer, S.; Eberle, T.; Edgar, M.; Edwards, M.; Effler, A.; Ehrens, P.; Endrőczi, G.; Engel, R.; Etzel, T.; Evans, K.; Evans, M.; Evans, T.; Factourovich, M.; Fafone, V.; Fairhurst, S.; Farr, B. F.; Farr, W. M.; Favata, M.; Fazi, D.; Fehrmann, H.; Feldbaum, D.; Feroz, F.; Ferrante, I.; Ferrini, F.; Fidecaro, F.; Finn, L. S.; Fiori, I.; Fisher, R. P.; Flaminio, R.; Foley, S.; Forsi, E.; Forte, L. A.; Fotopoulos, N.; Fournier, J.-D.; Franc, J.; Franco, S.; Frasca, S.; Frasconi, F.; Frede, M.; Frei, M. A.; Frei, Z.; Freise, A.; Frey, R.; Fricke, T. T.; Friedrich, D.; Fritschel, P.; Frolov, V. V.; Fujimoto, M.-K.; Fulda, P. J.; Fyffe, M.; Gair, J.; Galimberti, M.; Gammaitoni, L.; Garcia, J.; Garufi, F.; Gáspár, M. E.; Gelencser, G.; Gemme, G.; Genin, E.; Gennai, A.; Gergely, L. Á.; Ghosh, S.; Giaime, J. A.; Giampanis, S.; Giardina, K. D.; Giazotto, A.; Gil-Casanova, S.; Gill, C.; Gleason, J.; Goetz, E.; González, G.; Gorodetsky, M. L.; Goßler, S.; Gouaty, R.; Graef, C.; Graff, P. B.; Granata, M.; Grant, A.; Gray, C.; Greenhalgh, R. J. S.; Gretarsson, A. M.; Griffo, C.; Grote, H.; Grover, K.; Grunewald, S.; Guidi, G. M.; Guido, C.; Gupta, R.; Gustafson, E. K.; Gustafson, R.; Hallam, J. M.; Hammer, D.; Hammond, G.; Hanks, J.; Hanna, C.; Hanson, J.; Harms, J.; Harry, G. M.; Harry, I. W.; Harstad, E. D.; Hartman, M. T.; Haster, C.-J.; Haughian, K.; Hayama, K.; Hayau, J.-F.; Heefner, J.; Heidmann, A.; Heintze, M. C.; Heitmann, H.; Hello, P.; Hemming, G.; Hendry, M. A.; Heng, I. S.; Heptonstall, A. W.; Herrera, V.; Heurs, M.; Hewitson, M.; Hild, S.; Hoak, D.; Hodge, K. A.; Holt, K.; Holtrop, M.; Hong, T.; Hooper, S.; Hough, J.; Howell, E. J.; Hughey, B.; Husa, S.; Huttner, S. H.; Huynh-Dinh, T.; Ingram, D. R.; Inta, R.; Isogai, T.; Ivanov, A.; Izumi, K.; Jacobson, M.; James, E.; Jang, Y. J.; Jaranowski, P.; Jesse, E.; Johnson, W. W.; Jones, D. I.; Jones, R.; Jonker, R. J. G.; Ju, L.

    2013-09-01

    Compact binary systems with neutron stars or black holes are one of the most promising sources for ground-based gravitational-wave detectors. Gravitational radiation encodes rich information about source physics; thus parameter estimation and model selection are crucial analysis steps for any detection candidate events. Detailed models of the anticipated waveforms enable inference on several parameters, such as component masses, spins, sky location and distance, that are essential for new astrophysical studies of these sources. However, accurate measurements of these parameters and discrimination of models describing the underlying physics are complicated by artifacts in the data, uncertainties in the waveform models and in the calibration of the detectors. Here we report such measurements on a selection of simulated signals added either in hardware or software to the data collected by the two LIGO instruments and the Virgo detector during their most recent joint science run, including a “blind injection” where the signal was not initially revealed to the collaboration. We exemplify the ability to extract information about the source physics on signals that cover the neutron-star and black-hole binary parameter space over the component mass range 1M⊙-25M⊙ and the full range of spin parameters. The cases reported in this study provide a snapshot of the status of parameter estimation in preparation for the operation of advanced detectors.

  14. Binary Neutron Stars with Generic Spin, Eccentricity, Mass ratio, and Compactness - Quasi-equilibrium Sequences and First Evolutions

    CERN Document Server

    Dietrich, Tim; Johnson-McDaniel, Nathan K; Bernuzzi, Sebastiano; Markakis, Charalampos M; Bruegmann, Bernd; Tichy, Wolfgang

    2015-01-01

    Information about the last stages of a binary neutron star inspiral and the final merger can be extracted from quasi-equilibrium configurations and dynamical evolutions. In this article, we construct quasi-equilibrium configurations for different spins, eccentricities, mass ratios, compactnesses, and equations of state. For this purpose we employ the SGRID code, which allows us to construct such data in previously inaccessible regions of the parameter space. In particular, we consider spinning neutron stars in isolation and in binary systems; we incorporate new methods to produce highly eccentric and eccentricity reduced data; we present the possibility of computing data for significantly unequal-mass binaries; and we create equal-mass binaries with individual compactness up to 0.23. As a proof of principle, we explore the dynamical evolution of three new configurations. First, we simulate a $q=2.06$ mass ratio which is the highest mass ratio for a binary neutron star evolved in numerical relativity to date. ...

  15. Efficient generation of k-directional assembly sequences

    Energy Technology Data Exchange (ETDEWEB)

    Agarwal, P.K. [Duke Univ., Durham, NC (United States); Berg, M. de [Utrecht Univ. (Netherlands); Halperin, D. [Stanford Univ., CA (United States); Sharir, M. [Tel Aviv Univ. (Israel)

    1996-12-31

    Let S be a collection of n rigid bodies in 3-space, and let D be a set of k directions in 3-space, where k is a small constant. A k-directional assembly sequence for S, with respect to D, is a linear ordering (s{sub 1},...., s{sub n}) of the bodies in S, such that each si can be moved to infinity by translating it in one of the directions of D and without intersecting any s{sub j}, for j > i. We present an algorithm that computes a k-directional assembly sequence, or decides that no such sequence exists, for a set of polyhedra. The algorithm runs in O(km{sup 4/3+{epsilon}}) time, where m is the total number of vertices of the polyhedra. We also give an algorithm for {open_quote}k-directional{close_quote} rotational motions.

  16. Generating long sequences of high-intensity femtosecond pulses

    CERN Document Server

    Bitter, Martin

    2015-01-01

    We present an approach to create pulse sequences extending beyond 150~picoseconds in duration, comprised of $100~\\mu$J femtosecond pulses. A quarter of the pulse train is produced by a high-resolution pulse shaper, which allows full controllability over the timing of each pulse. Two nested Michelson interferometers follow to quadruple the pulse number and the sequence duration. To boost the pulse energy, the long train is sent through a multi-pass Ti:Sapphire amplifier, followed by an external compressor. A periodic sequence of 84~pulses of 120~fs width and an average pulse energy of 107~$\\mu$J, separated by 2~ps, is demonstrated as a proof of principle.

  17. STATE SPACE GENERATION FRAMEWORK BASED ON BINARY DECISION DIAGRAM FOR DISTRIBUTED EXPLICIT MODEL CHECKING

    Directory of Open Access Journals (Sweden)

    Nacer Tabib

    2016-01-01

    Full Text Available This paper proposes a new framework based on Binary Decision Diagrams (BDD for the graph distribution problem in the context of explicit model checking. The BDD are yet used to represent the state space for a symbolic verification model checking. Thus, we took advantage of high compression ratio of BDD to encode not only the state space, but also the place where each state will be put. So, a fitness function that allows a good balance load of states over the nodes of an homogeneous network is used. Furthermore, a detailed explanation of how to calculate the inter-site edges between different nodes based on the adapted data structure is presented.

  18. Parameter estimation for compact binary coalescence signals with the first generation gravitational-wave detector network

    CERN Document Server

    Aasi, J; Abbott, B P; Abbott, R; Abbott, T D; Abernathy, M; Accadia, T; Acernese, F; Adams, C; Adams, T; Addesso, P; Adhikari, R; Affeldt, C; Agathos, M; Agatsuma, K; Ajith, P; Allen, B; Allocca, A; Ceron, E Amador; Amariutei, D; Anderson, S B; Anderson, W G; Arai, K; Araya, M C; Ast, S; Aston, S M; Astone, P; Atkinson, D; Aufmuth, P; Aulbert, C; Aylott, B E; Babak, S; Baker, P; Ballardin, G; Ballmer, S; Bao, Y; Barayoga, J C B; Barker, D; Barone, F; Barr, B; Barsotti, L; Barsuglia, M; Barton, M A; Bartos, I; Bassiri, R; Bastarrika, M; Basti, A; Batch, J; Bauchrowitz, J; Bauer, Th S; Bebronne, M; Beck, D; Behnke, B; Bejger, M; Beker, M G; Bell, A S; Bell, C; Belopolski, I; Benacquista, M; Berliner, J M; Bertolini, A; Betzwieser, J; Beveridge, N; Beyersdorf, P T; Bhadbade, T; Bilenko, I A; Billingsley, G; Birch, J; Biswas, R; Bitossi, M; Bizouard, M A; Black, E; Blackburn, J K; Blackburn, L; Blair, D; Bland, B; Blom, M; Bock, O; Bodiya, T P; Bogan, C; Bond, C; Bondarescu, R; Bondu, F; Bonelli, L; Bonnand, R; Bork, R; Born, M; Boschi, V; Bose, S; Bosi, L; Bouhou, B; Braccini, S; Bradaschia, C; Brady, P R; Braginsky, V B; Branchesi, M; Brau, J E; Breyer, J; Briant, T; Bridges, D O; Brillet, A; Brinkmann, M; Brisson, V; Britzger, M; Brooks, A F; Brown, D A; Bulik, T; Bulten, H J; Buonanno, A; Burguet--Castell, J; Buskulic, D; Buy, C; Byer, R L; Cadonati, L; Cagnoli, G; Calloni, E; Camp, J B; Campsie, P; Cannon, K; Canuel, B; Cao, J; Capano, C D; Carbognani, F; Carbone, L; Caride, S; Caudill, S; Cavaglià, M; Cavalier, F; Cavalieri, R; Cella, G; Cepeda, C; Cesarini, E; Chalermsongsak, T; Charlton, P; Chassande-Mottin, E; Chen, W; Chen, X; Chen, Y; Chincarini, A; Chiummo, A; Cho, H S; Chow, J; Christensen, N; Chua, S S Y; Chung, C T Y; Chung, S; Ciani, G; Clara, F; Clark, D E; Clark, J A; Clayton, J H; Cleva, F; Coccia, E; Cohadon, P -F; Colacino, C N; Colla, A; Colombini, M; Conte, A; Conte, R; Cook, D; Corbitt, T R; Cordier, M; Cornish, N; Corsi, A; Costa, C A; Coughlin, M; Coulon, J -P; Couvares, P; Coward, D M; Cowart, M; Coyne, D C; Creighton, J D E; Creighton, T D; Cruise, A M; Cumming, A; Cunningham, L; Cuoco, E; Cutler, R M; Dahl, K; Damjanic, M; Danilishin, S L; D'Antonio, S; Danzmann, K; Dattilo, V; Daudert, B; Daveloza, H; Davier, M; Daw, E J; Dayanga, T; De Rosa, R; DeBra, D; Debreczeni, G; Degallaix, J; Del Pozzo, W; Dent, T; Dergachev, V; DeRosa, R; Dhurandhar, S; Di Fiore, L; Di Lieto, A; Di Palma, I; Emilio, M Di Paolo; Di Virgilio, A; Díaz, M; Dietz, A; Donovan, F; Dooley, K L; Doravari, S; Dorsher, S; Drago, M; Drever, R W P; Driggers, J C; Du, Z; Dumas, J -C; Dwyer, S; Eberle, T; Edgar, M; Edwards, M; Effler, A; Ehrens, P; Endröczi, G; Engel, R; Etzel, T; Evans, K; Evans, M; Evans, T; Factourovich, M; Fafone, V; Fairhurst, S; Farr, B F; Farr, W M; Favata, M; Fazi, D; Fehrmann, H; Feldbaum, D; Ferrante, I; Ferrini, F; Fidecaro, F; Finn, L S; Fiori, I; Fisher, R P; Flaminio, R; Foley, S; Forsi, E; Forte, L A; Fotopoulos, N; Fournier, J -D; Franc, J; Franco, S; Frasca, S; Frasconi, F; Frede, M; Frei, M A; Frei, Z; Freise, A; Frey, R; Fricke, T T; Friedrich, D; Fritschel, P; Frolov, V V; Fujimoto, M -K; Fulda, P J; Fyffe, M; Gair, J; Galimberti, M; Gammaitoni, L; Garcia, J; Garufi, F; Gáspár, M E; Gelencser, G; Gemme, G; Genin, E; Gennai, A; Gergely, L Á; Ghosh, S; Giaime, J A; Giampanis, S; Giardina, K D; Giazotto, A; Gil-Casanova, S; Gill, C; Gleason, J; Goetz, E; González, G; Gorodetsky, M L; Goßler, S; Gouaty, R; Graef, C; Graff, P B; Granata, M; Grant, A; Gray, C; Greenhalgh, R J S; Gretarsson, A M; Griffo, C; Grote, H; Grover, K; Grunewald, S; Guidi, G M; Guido, C; Gupta, R; Gustafson, E K; Gustafson, R; Hallam, J M; Hammer, D; Hammond, G; Hanks, J; Hanna, C; Hanson, J; Harms, J; Harry, G M; Harry, I W; Harstad, E D; Hartman, M T; Haster, C -J; Haughian, K; Hayama, K; Hayau, J -F; Heefner, J; Heidmann, A; Heintze, M C; Heitmann, H; Hello, P; Hemming, G; Hendry, M A; Heng, I S; Heptonstall, A W; Herrera, V; Heurs, M; Hewitson, M; Hild, S; Hoak, D; Hodge, K A; Holt, K; Holtrop, M; Hong, T; Hooper, S; Hough, J; Howell, E J; Hughey, B; Husa, S; Huttner, S H; Huynh-Dinh, T; Ingram, D R; Inta, R; Isogai, T; Ivanov, A; Izumi, K; Jacobson, M; James, E; Jang, Y J; Jaranowski, P; Jesse, E; Johnson, W W; Jones, D I; Jones, R; Jonker, R J G; Ju, L; Kalmus, P; Kalogera, V; Kandhasamy, S; Kang, G; Kanner, J B; Kasprzack, M; Kasturi, R; Katsavounidis, E; Katzman, W; Kaufer, H; Kaufman, K; Kawabe, K; Kawamura, S; Kawazoe, F; Keitel, D; Kelley, D; Kells, W; Keppel, D G; Keresztes, Z; Khalaidovski, A; Khalili, F Y; Khazanov, E A; Kim, B K; Kim, C; Kim, H; Kim, K; Kim, N; Kim, Y M; King, P J; Kinzel, D L; Kissel, J S; Klimenko, S; Kline, J; Kokeyama, K; Kondrashov, V; Koranda, S; Korth, W Z; Kowalska, I; Kozak, D; Kringel, V; Krishnan, B; Królak, A; Kuehn, G; Kumar, P; Kumar, R; Kurdyumov, R; Kwee, P; Lam, P K; Landry, M; Langley, A; Lantz, B; Lastzka, N; Lawrie, C; Lazzarini, A; Roux, A Le; Leaci, P; Lee, C H; Lee, H K; Lee, H M; Leong, J R; Leonor, I; Leroy, N; Letendre, N; Lhuillier, V; Li, J; Li, T G F; Lindquist, P E; Litvine, V; Liu, Y; Liu, Z; Lockerbie, N A; Lodhia, D; Logue, J; Lorenzini, M; Loriette, V; Lormand, M; Losurdo, G; Lough, J; Lubinski, M; Lück, H; Lundgren, A P; Macarthur, J; Macdonald, E; Machenschalk, B; MacInnis, M; Macleod, D M; Mageswaran, M; Mailand, K; Majorana, E; Maksimovic, I; Malvezzi, V; Man, N; Mandel, I; Mandic, V; Mantovani, M; Marchesoni, F; Marion, F; Márka, S; Márka, Z; Markosyan, A; Maros, E; Marque, J; Martelli, F; Martin, I W; Martin, R M; Marx, J N; Mason, K; Masserot, A; Matichard, F; Matone, L; Matzner, R A; Mavalvala, N; Mazzolo, G; McCarthy, R; McClelland, D E; McGuire, S C; McIntyre, G; McIver, J; Meadors, G D; Mehmet, M; Meier, T; Melatos, A; Melissinos, A C; Mendell, G; Menéndez, D F; Mercer, R A; Meshkov, S; Messenger, C; Meyer, M S; Miao, H; Michel, C; Milano, L; Miller, J; Minenkov, Y; Mingarelli, C M F; Mitrofanov, V P; Mitselmakher, G; Mittleman, R; Moe, B; Mohan, M; Mohapatra, S R P; Moraru, D; Moreno, G; Morgado, N; Morgia, A; Mori, T; Morriss, S R; Mosca, S; Mossavi, K; Mours, B; Mow--Lowry, C M; Mueller, C L; Mueller, G; Mukherjee, S; Mullavey, A; Müller-Ebhardt, H; Munch, J; Murphy, D; Murray, P G; Mytidis, A; Nash, T; Naticchioni, L; Necula, V; Nelson, J; Neri, I; Newton, G; Nguyen, T; Nishizawa, A; Nitz, A; Nocera, F; Nolting, D; Normandin, M E; Nuttall, L; Ochsner, E; O'Dell, J; Oelker, E; Ogin, G H; Oh, J J; Oh, S H; Oldenberg, R G; O'Reilly, B; O'Shaughnessy, R; Osthelder, C; Ott, C D; Ottaway, D J; Ottens, R S; Overmier, H; Owen, B J; Page, A; Palladino, L; Palomba, C; Pan, Y; Pankow, C; Paoletti, F; Paoletti, R; Papa, M A; Parisi, M; Pasqualetti, A; Passaquieti, R; Passuello, D; Pedraza, M; Penn, S; Perreca, A; Persichetti, G; Phelps, M; Pichot, M; Pickenpack, M; Piergiovanni, F; Pierro, V; Pihlaja, M; Pinard, L; Pinto, I M; Pitkin, M; Pletsch, H J; Plissi, M V; Poggiani, R; Pöld, J; Postiglione, F; Poux, C; Prato, M; Predoi, V; Prestegard, T; Price, L R; Prijatelj, M; Principe, M; Privitera, S; Prodi, G A; Prokhorov, L G; Puncken, O; Punturo, M; Puppo, P; Quetschke, V; Quitzow-James, R; Raab, F J; Rabeling, D S; Rácz, I; Radkins, H; Raffai, P; Rakhmanov, M; Ramet, C; Rankins, B; Rapagnani, P; Raymond, V; Re, V; Reed, C M; Reed, T; Regimbau, T; Reid, S; Reitze, D H; Ricci, F; Riesen, R; Riles, K; Roberts, M; Robertson, N A; Robinet, F; Robinson, C; Robinson, E L; Rocchi, A; Roddy, S; Rodriguez, C; Rodruck, M; Rolland, L; Rollins, J G; Romano, R; Romie, J H; Rosińska, D; Röver, C; Rowan, S; Rüdiger, A; Ruggi, P; Ryan, K; Salemi, F; Sammut, L; Sandberg, V; Sankar, S; Sannibale, V; Santamaría, L; Santiago-Prieto, I; Santostasi, G; Saracco, E; Sassolas, B; Sathyaprakash, B S; Saulson, P R; Savage, R L; Schilling, R; Schnabel, R; Schofield, R M S; Schulz, B; Schutz, B F; Schwinberg, P; Scott, J; Scott, S M; Seifert, F; Sellers, D; Sentenac, D; Sergeev, A; Shaddock, D A; Shaltev, M; Shapiro, B; Shawhan, P; Shoemaker, D H; Sidery, T L; Siemens, X; Sigg, D; Simakov, D; Singer, A; Singer, L; Sintes, A M; Skelton, G R; Slagmolen, B J J; Slutsky, J; Smith, J R; Smith, M R; Smith, R J E; Smith-Lefebvre, N D; Somiya, K; Sorazu, B; Speirits, F C; Sperandio, L; Stefszky, M; Steinert, E; Steinlechner, J; Steinlechner, S; Steplewski, S; Stochino, A; Stone, R; Strain, K A; Strigin, S E; Stroeer, A S; Sturani, R; Stuver, A L; Summerscales, T Z; Sung, M; Susmithan, S; Sutton, P J; Swinkels, B; Szeifert, G; Tacca, M; Taffarello, L; Talukder, D; Tanner, D B; Tarabrin, S P; Taylor, R; ter Braack, A P M; Thomas, P; Thorne, K A; Thorne, K S; Thrane, E; Thüring, A; Titsler, C; Tokmakov, K V; Tomlinson, C; Toncelli, A; Tonelli, M; Torre, O; Torres, C V; Torrie, C I; Tournefier, E; Travasso, F; Traylor, G; Tse, M; Ugolini, D; Vahlbruch, H; Vajente, G; Brand, J F J van den; Broeck, C Van Den; van der Putten, S; van Veggel, A A; Vass, S; Vasuth, M; Vaulin, R; Vavoulidis, M; Vecchio, A; Vedovato, G; Veitch, J; Veitch, P J; Venkateswara, K; Verkindt, D; Vetrano, F; Viceré, A; Villar, A E; Vinet, J -Y; Vitale, S; Vocca, H; Vorvick, C; Vyatchanin, S P; Wade, A; Wade, L; Wade, M; Waldman, S J; Wallace, L; Wan, Y; Wang, M; Wang, X; Wanner, A; Ward, R L; Was, M; Weinert, M; Weinstein, A J; Weiss, R; Welborn, T; Wen, L; Wessels, P; West, M; Westphal, T; Wette, K; Whelan, J T; Whitcomb, S E; White, D J; Whiting, B F; Wiesner, K; Wilkinson, C; Willems, P A; Williams, L; Williams, R; Willke, B; Wimmer, M; Winkelmann, L; Winkler, W; Wipf, C C; Wiseman, A G; Wittel, H; Woan, G; Wooley, R; Worden, J; Yablon, J; Yakushin, I; Yamamoto, H; Yamamoto, K; Yancey, C C; Yang, H; Yeaton-Massey, D; Yoshida, S; Yvert, M; Zadrożny, A; Zanolin, M; Zendri, J -P; Zhang, F; Zhang, L; Zhao, C; Zotov, N; Zucker, M E; Zweizig, J

    2013-01-01

    Compact binary systems with neutron stars or black holes are one of the most promising sources for ground-based gravitational wave detectors. Gravitational radiation encodes rich information about source physics; thus parameter estimation and model selection are crucial analysis steps for any detection candidate events. Detailed models of the anticipated waveforms enable inference on several parameters, such as component masses, spins, sky location and distance that are essential for new astrophysical studies of these sources. However, accurate measurements of these parameters and discrimination of models describing the underlying physics are complicated by artifacts in the data, uncertainties in the waveform models and in the calibration of the detectors. Here we report such measurements on a selection of simulated signals added either in hardware or software to the data collected by the two LIGO instruments and the Virgo detector during their most recent joint science run, including a "blind injection" wher...

  19. A survey of sequence alignment algorithms for next-generation sequencing.

    Science.gov (United States)

    Li, Heng; Homer, Nils

    2010-09-01

    Rapidly evolving sequencing technologies produce data on an unparalleled scale. A central challenge to the analysis of this data is sequence alignment, whereby sequence reads must be compared to a reference. A wide variety of alignment algorithms and software have been subsequently developed over the past two years. In this article, we will systematically review the current development of these algorithms and introduce their practical applications on different types of experimental data. We come to the conclusion that short-read alignment is no longer the bottleneck of data analyses. We also consider future development of alignment algorithms with respect to emerging long sequence reads and the prospect of cloud computing.

  20. Applications and Case Studies of the Next-Generation Sequencing Technologies in Food, Nutrition and Agriculture.

    Science.gov (United States)

    Next-generation sequencing technologies are able to produce high-throughput short sequence reads in a cost-effective fashion. The emergence of these technologies has not only facilitated genome sequencing but also changed the landscape of life sciences. Here I survey their major applications ranging...

  1. Application of next generation sequencing in clinical microbiology and infection prevention

    NARCIS (Netherlands)

    Deurenberg, Ruud H; Bathoorn, Erik; Chlebowicz, Monika A; Couto, Natacha; Ferdous, Mithila; García-Cobos, Silvia; Kooistra-Smid, Anna M D; Raangs, Erwin C; Rosema, Sigrid; Veloo, Alida C M; Zhou, Kai; Friedrich, Alexander W; Rossen, John W A

    2016-01-01

    Current molecular diagnostics of human pathogens provide limited information that is often not sufficient for outbreak and transmission investigation. Next generation sequencing (NGS) determines the DNA sequence of a complete bacterial genome in a single sequence run, and from these data, informatio

  2. GRASS: a generic algorithm for scaffolding next-generation sequencing assemblies

    NARCIS (Netherlands)

    Gritsenko, A.A.; Nijkamp, J.F.; Reinders, M.J.T.; De Ridder, D.

    2012-01-01

    Motivation: The increasing availability of second-generation highthroughput sequencing (HTS) technologies has sparked a growing interest in de novo genome sequencing. This in turn has fueled the need for reliable means of obtaining high-quality draft genomes from short-read sequencing data. The mill

  3. A video annotation methodology for interactive video sequence generation

    NARCIS (Netherlands)

    Lindley, C.A.; Earnshaw, R.A.; Vince, J.A.

    2001-01-01

    The FRAMES project within the RDN CRC (Cooperative Research Centre for Research Data Networks) has developed an experimental environment for dynamic virtual video sequence synthesis from databases of video data. A major issue for the development of dynamic interactive video applications of this type

  4. Next generation sequencing and its applications in forensic genetics

    DEFF Research Database (Denmark)

    Børsting, Claus; Morling, Niels

    2015-01-01

    matured during the last 10 years, and the quality of the sequences has reached a level where NGS is used in clinical diagnostics of humans. Forensic genetic laboratories have also explored NGS technologies and especially in the last year, there has been a small explosion in the number of scientific...... articles and presentations at conferences with forensic aspects of NGS. These contributions have demonstrated that NGS offers new possibilities for forensic genetic case work. More information may be obtained from unique samples in a single experiment by analyzing combinations of markers (STRs, SNPs...... and will increase the statistical weight of the evidence. In this review, we will give an introduction to NGS and single-molecule sequencing, and we will discuss the possible applications of NGS in forensic genetics....

  5. Harnessing Next-Generation Sequencing Capabilities for Microbial Forensics

    Science.gov (United States)

    2014-07-15

    a sophisticated analysis identified the source, many countries acted proactively to remove cucumbers and tomatoes from store shelves, Russia issued...Eisenstein, M. (2012). Oxford Nanopore announcement sets sequencing sector abuzz. Nature Biotechnology , 30(4), 295–6. doi:10.1038/nbt0412-295... Biotechnology , 29(7). Grad, Y. H., Lipsitch, M., Feldgarden, M., Arachchi, H. M., Cerqueira, G. C., Fitzgerald, M., .. . Hanage, W. P. (2012). Genomic

  6. Identification of optimum sequencing depth especially for de novo genome assembly of small genomes using next generation sequencing data.

    Science.gov (United States)

    Desai, Aarti; Marwah, Veer Singh; Yadav, Akshay; Jha, Vineet; Dhaygude, Kishor; Bangar, Ujwala; Kulkarni, Vivek; Jere, Abhay

    2013-01-01

    Next Generation Sequencing (NGS) is a disruptive technology that has found widespread acceptance in the life sciences research community. The high throughput and low cost of sequencing has encouraged researchers to undertake ambitious genomic projects, especially in de novo genome sequencing. Currently, NGS systems generate sequence data as short reads and de novo genome assembly using these short reads is computationally very intensive. Due to lower cost of sequencing and higher throughput, NGS systems now provide the ability to sequence genomes at high depth. However, currently no report is available highlighting the impact of high sequence depth on genome assembly using real data sets and multiple assembly algorithms. Recently, some studies have evaluated the impact of sequence coverage, error rate and average read length on genome assembly using multiple assembly algorithms, however, these evaluations were performed using simulated datasets. One limitation of using simulated datasets is that variables such as error rates, read length and coverage which are known to impact genome assembly are carefully controlled. Hence, this study was undertaken to identify the minimum depth of sequencing required for de novo assembly for different sized genomes using graph based assembly algorithms and real datasets. Illumina reads for E.coli (4.6 MB) S.kudriavzevii (11.18 MB) and C.elegans (100 MB) were assembled using SOAPdenovo, Velvet, ABySS, Meraculous and IDBA-UD. Our analysis shows that 50X is the optimum read depth for assembling these genomes using all assemblers except Meraculous which requires 100X read depth. Moreover, our analysis shows that de novo assembly from 50X read data requires only 6-40 GB RAM depending on the genome size and assembly algorithm used. We believe that this information can be extremely valuable for researchers in designing experiments and multiplexing which will enable optimum utilization of sequencing as well as analysis resources.

  7. Three observational differences for binary black holes detections with second and third generation gravitational-wave detectors

    CERN Document Server

    Vitale, Salvatore

    2016-01-01

    Advanced gravitational-wave observatories, such as LIGO and Virgo, will detect hundreds of gravitational waves emitted by binary black holes in the next few years. The collection of detected sources is expected to have certain properties. It is expected that a selection bias will exist toward higher mass systems, that most events will be oriented with their angular momentum pointing to or away from Earth, and that quiet events will be much more numerous than loud events. In this paper we show how all these assumptions are only true for existing detectors and do not have any universality. Using an network of proposed third-generation gravitational wave detectors, we show how each of these assumptions must be revised and we discuss several consequences on the characterization of the sources.

  8. Three observational differences for binary black holes detections with second- and third-generation gravitational-wave detectors

    Science.gov (United States)

    Vitale, Salvatore

    2016-12-01

    Advanced gravitational-wave observatories, such as LIGO and Virgo, will detect hundreds of gravitational-wave signals emitted by binary black holes in the next few years. The collection of detected sources is expected to have certain properties. It is expected that a selection bias will exist toward higher-mass systems, that most events will be oriented with their angular momentum pointing to or away from Earth, and that quiet events will be much more numerous than loud events. In this paper, we show how all these assumptions are only true for existing detectors and do not have any universality. Using a network of proposed third-generation gravitational-wave detectors, we show how each of these assumptions must be revised, and we discuss several consequences on the characterization of the sources.

  9. Surface behaviour and undercooling in the liquid Ga-Bi binary system detected by optical second harmonic generation

    Institute of Scientific and Technical Information of China (English)

    王聪; 王天民

    2003-01-01

    We employ an optical second harmonic generation(SHG) technique to investigate the surfeace behaviours at the liquid(solid)/vapour interface of the Ga-Bi binary metallic system. In a heating and cooling cycle between 280℃ and room temperature, there is no change of the SH-intensity in the heating process, whereas there exists an abrupt and abnormal change of the SH-intensity in the cooling process. It is interesting to find that a macroscopic Bi-rich solid layer is floating on the surface of the Ga-rich liquid phase just below the monotectic temperature (222℃±2℃) in the cooling process, in spite of the Bi-rich phase being heavier than the Ga-rich phase. On the other hand, different undercooling behaviours are observed at the surface and in the bulk. The behaviours of surface solidification and surface melting are different from those in the bulk.

  10. Using binary optical elements (BOEs) to generate rectangular spots for illumination in micro flow cytometer

    Science.gov (United States)

    Zhao, Jingjing; You, Zheng

    2016-01-01

    This work introduces three rectangular quasi-flat-top spots, which are provided by binary optical elements (BOEs) and utilized for the illumination in a microflow cytometer. The three spots contain, respectively, one, two, and three rectangles (R1, R2, and R3). To test the performance of this mechanism, a microflow cytometer is established by integrating the BOEs and a three-dimensional hydrodynamic focusing chip. Through the experiments of detecting fluorescence microbeads, the three spots present good fluorescence coefficients of variation in comparison with those derived from commercial instruments. Benefiting from a high spatial resolution, when using R1 spot, the micro flow cytometer can perform a throughput as high as 20 000 events per second (eps). Illuminated by R2 or R3 spot, one bead emits fluorescence twice or thrice, thus the velocity can be measured in real time. Besides, the R3 spot provides a long-time exposure, which is conducive to improving fluorescence intensity and the measurement stability. In brief, using the spots shaped and homogenized by BOEs for illumination can increase the performance and the functionality of a micro flow cytometer. PMID:27733892

  11. Quantifying Next Generation Sequencing Sample Pre-Processing Bias in HIV-1 Complete Genome Sequencing.

    Science.gov (United States)

    Vrancken, Bram; Trovão, Nídia Sequeira; Baele, Guy; van Wijngaerden, Eric; Vandamme, Anne-Mieke; van Laethem, Kristel; Lemey, Philippe

    2016-01-07

    Genetic analyses play a central role in infectious disease research. Massively parallelized "mechanical cloning" and sequencing technologies were quickly adopted by HIV researchers in order to broaden the understanding of the clinical importance of minor drug-resistant variants. These efforts have, however, remained largely limited to small genomic regions. The growing need to monitor multiple genome regions for drug resistance testing, as well as the obvious benefit for studying evolutionary and epidemic processes makes complete genome sequencing an important goal in viral research. In addition, a major drawback for NGS applications to RNA viruses is the need for large quantities of input DNA. Here, we use a generic overlapping amplicon-based near full-genome amplification protocol to compare low-input enzymatic fragmentation (Nextera™) with conventional mechanical shearing for Roche 454 sequencing. We find that the fragmentation method has only a modest impact on the characterization of the population composition and that for reliable results, the variation introduced at all steps of the procedure--from nucleic acid extraction to sequencing--should be taken into account, a finding that is also relevant for NGS technologies that are now more commonly used. Furthermore, by applying our protocol to deep sequence a number of pre-therapy plasma and PBMC samples, we illustrate the potential benefits of a near complete genome sequencing approach in routine genotyping.

  12. Exact distribution of a pattern in a set of random sequences generated by a Markov source: applications to biological data

    Directory of Open Access Journals (Sweden)

    Regad Leslie

    2010-01-01

    Full Text Available Abstract Background In bioinformatics it is common to search for a pattern of interest in a potentially large set of rather short sequences (upstream gene regions, proteins, exons, etc.. Although many methodological approaches allow practitioners to compute the distribution of a pattern count in a random sequence generated by a Markov source, no specific developments have taken into account the counting of occurrences in a set of independent sequences. We aim to address this problem by deriving efficient approaches and algorithms to perform these computations both for low and high complexity patterns in the framework of homogeneous or heterogeneous Markov models. Results The latest advances in the field allowed us to use a technique of optimal Markov chain embedding based on deterministic finite automata to introduce three innovative algorithms. Algorithm 1 is the only one able to deal with heterogeneous models. It also permits to avoid any product of convolution of the pattern distribution in individual sequences. When working with homogeneous models, Algorithm 2 yields a dramatic reduction in the complexity by taking advantage of previous computations to obtain moment generating functions efficiently. In the particular case of low or moderate complexity patterns, Algorithm 3 exploits power computation and binary decomposition to further reduce the time complexity to a logarithmic scale. All these algorithms and their relative interest in comparison with existing ones were then tested and discussed on a toy-example and three biological data sets: structural patterns in protein loop structures, PROSITE signatures in a bacterial proteome, and transcription factors in upstream gene regions. On these data sets, we also compared our exact approaches to the tempting approximation that consists in concatenating the sequences in the data set into a single sequence. Conclusions Our algorithms prove to be effective and able to handle real data sets with

  13. Challenges and opportunities for next-generation sequencing in companion diagnostics.

    Science.gov (United States)

    Lin, Erick; Chien, Jeremy; Ong, Frank S; Fan, Jian-Bing

    2015-02-01

    The rapid decline in sequencing costs has allowed next-generation sequencing (NGS) assays, previously ubiquitous only in research laboratories, to begin making inroads into molecular diagnostics. Genotypic assays - DNA sequencing - include whole genome sequencing, whole exome sequencing, focused assays that target only a handful of genes. Phenotypic assays comprise a broader spectrum of options and can query a variety of epigenetic modifications of DNA (such as ChIP-seq, bisulfite sequencing, DNase-I hypersensitivity site-sequencing, Formaldehyde-Assisted Isolation of Regulatory Elements-sequencing, etc.) that regulate gene expression-related processes or gene expression (RNA-sequencing) itself. To date, the US FDA has only cleared 12 DNA-based companion diagnostic tests, all in cancer. Although challenges exist for NGS in companion diagnostics, the wide-ranging capabilities of NGS offer extraordinary opportunities for the development and implementation of NGS-based companion diagnostics to probe oncogenes, tumor suppressor genes and cancer-enabling genes.

  14. A tale of three next generation sequencing platforms: comparison of Ion Torrent, Pacific Biosciences and Illumina MiSeq sequencers

    Directory of Open Access Journals (Sweden)

    Quail Michael A

    2012-07-01

    Full Text Available Abstract Background Next generation sequencing (NGS technology has revolutionized genomic and genetic research. The pace of change in this area is rapid with three major new sequencing platforms having been released in 2011: Ion Torrent’s PGM, Pacific Biosciences’ RS and the Illumina MiSeq. Here we compare the results obtained with those platforms to the performance of the Illumina HiSeq, the current market leader. In order to compare these platforms, and get sufficient coverage depth to allow meaningful analysis, we have sequenced a set of 4 microbial genomes with mean GC content ranging from 19.3 to 67.7%. Together, these represent a comprehensive range of genome content. Here we report our analysis of that sequence data in terms of coverage distribution, bias, GC distribution, variant detection and accuracy. Results Sequence generated by Ion Torrent, MiSeq and Pacific Biosciences technologies displays near perfect coverage behaviour on GC-rich, neutral and moderately AT-rich genomes, but a profound bias was observed upon sequencing the extremely AT-rich genome of Plasmodium falciparum on the PGM, resulting in no coverage for approximately 30% of the genome. We analysed the ability to call variants from each platform and found that we could call slightly more variants from Ion Torrent data compared to MiSeq data, but at the expense of a higher false positive rate. Variant calling from Pacific Biosciences data was possible but higher coverage depth was required. Context specific errors were observed in both PGM and MiSeq data, but not in that from the Pacific Biosciences platform. Conclusions All three fast turnaround sequencers evaluated here were able to generate usable sequence. However there are key differences between the quality of that data and the applications it will support.

  15. Quantifying Next Generation Sequencing Sample Pre-Processing Bias in HIV-1 Complete Genome Sequencing

    Directory of Open Access Journals (Sweden)

    Bram Vrancken

    2016-01-01

    Full Text Available Genetic analyses play a central role in infectious disease research. Massively parallelized “mechanical cloning” and sequencing technologies were quickly adopted by HIV researchers in order to broaden the understanding of the clinical importance of minor drug-resistant variants. These efforts have, however, remained largely limited to small genomic regions. The growing need to monitor multiple genome regions for drug resistance testing, as well as the obvious benefit for studying evolutionary and epidemic processes makes complete genome sequencing an important goal in viral research. In addition, a major drawback for NGS applications to RNA viruses is the need for large quantities of input DNA. Here, we use a generic overlapping amplicon-based near full-genome amplification protocol to compare low-input enzymatic fragmentation (Nextera™ with conventional mechanical shearing for Roche 454 sequencing. We find that the fragmentation method has only a modest impact on the characterization of the population composition and that for reliable results, the variation introduced at all steps of the procedure—from nucleic acid extraction to sequencing—should be taken into account, a finding that is also relevant for NGS technologies that are now more commonly used. Furthermore, by applying our protocol to deep sequence a number of pre-therapy plasma and PBMC samples, we illustrate the potential benefits of a near complete genome sequencing approach in routine genotyping.

  16. An efficient quantitation method of next-generation sequencing libraries by using MiSeq sequencer.

    Science.gov (United States)

    Katsuoka, Fumiki; Yokozawa, Junji; Tsuda, Kaoru; Ito, Shin; Pan, Xiaoqing; Nagasaki, Masao; Yasuda, Jun; Yamamoto, Masayuki

    2014-12-01

    Library quantitation is a critical step to obtain high data output in Illumina HiSeq sequencers. Here, we introduce a library quantitation method that uses the Illumina MiSeq sequencer designated as quantitative MiSeq (qMiSeq). In this procedure, 96 dual-index libraries, including control samples, are denatured, pooled in equal volume, and sequenced by MiSeq. We found that relative concentration of each library can be determined based on the observed index ratio and can be used to determine HiSeq run condition for each library. Thus, qMiSeq provides an efficient way to quantitate a large number of libraries at a time.

  17. Next generation sequencing of DNA-launched Chikungunya vaccine virus

    Energy Technology Data Exchange (ETDEWEB)

    Hidajat, Rachmat; Nickols, Brian [Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701 (United States); Forrester, Naomi [Institute for Human Infections and Immunity, Sealy Center for Vaccine Development and Department of Pathology, University of Texas Medical Branch, GNL, 301 University Blvd., Galveston, TX 77555 (United States); Tretyakova, Irina [Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701 (United States); Weaver, Scott [Institute for Human Infections and Immunity, Sealy Center for Vaccine Development and Department of Pathology, University of Texas Medical Branch, GNL, 301 University Blvd., Galveston, TX 77555 (United States); Pushko, Peter, E-mail: ppushko@medigen-usa.com [Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD 21701 (United States)

    2016-03-15

    Chikungunya virus (CHIKV) represents a pandemic threat with no approved vaccine available. Recently, we described a novel vaccination strategy based on iDNA® infectious clone designed to launch a live-attenuated CHIKV vaccine from plasmid DNA in vitro or in vivo. As a proof of concept, we prepared iDNA plasmid pCHIKV-7 encoding the full-length cDNA of the 181/25 vaccine. The DNA-launched CHIKV-7 virus was prepared and compared to the 181/25 virus. Illumina HiSeq2000 sequencing revealed that with the exception of the 3′ untranslated region, CHIKV-7 viral RNA consistently showed a lower frequency of single-nucleotide polymorphisms than the 181/25 RNA including at the E2-12 and E2-82 residues previously identified as attenuating mutations. In the CHIKV-7, frequencies of reversions at E2-12 and E2-82 were 0.064% and 0.086%, while in the 181/25, frequencies were 0.179% and 0.133%, respectively. We conclude that the DNA-launched virus has a reduced probability of reversion mutations, thereby enhancing vaccine safety. - Highlights: • Chikungunya virus (CHIKV) is an emerging pandemic threat. • In vivo DNA-launched attenuated CHIKV is a novel vaccine technology. • DNA-launched virus was sequenced using HiSeq2000 and compared to the 181/25 virus. • DNA-launched virus has lower frequency of SNPs at E2-12 and E2-82 attenuation loci.

  18. From Conventional to Next Generation Sequencing of Epstein-Barr Virus Genomes.

    Science.gov (United States)

    Kwok, Hin; Chiang, Alan Kwok Shing

    2016-02-24

    Genomic sequences of Epstein-Barr virus (EBV) have been of interest because the virus is associated with cancers, such as nasopharyngeal carcinoma, and conditions such as infectious mononucleosis. The progress of whole-genome EBV sequencing has been limited by the inefficiency and cost of the first-generation sequencing technology. With the advancement of next-generation sequencing (NGS) and target enrichment strategies, increasing number of EBV genomes has been published. These genomes were sequenced using different approaches, either with or without EBV DNA enrichment. This review provides an overview of the EBV genomes published to date, and a description of the sequencing technology and bioinformatic analyses employed in generating these sequences. We further explored ways through which the quality of sequencing data can be improved, such as using DNA oligos for capture hybridization, and longer insert size and read length in the sequencing runs. These advances will enable large-scale genomic sequencing of EBV which will facilitate a better understanding of the genetic variations of EBV in different geographic regions and discovery of potentially pathogenic variants in specific diseases.

  19. Generating Multiple Base-Resolution DNA Methylomes Using Reduced Representation Bisulfite Sequencing.

    Science.gov (United States)

    Chatterjee, Aniruddha; Rodger, Euan J; Stockwell, Peter A; Le Mée, Gwenn; Morison, Ian M

    2017-01-01

    Reduced representation bisulfite sequencing (RRBS) is an effective technique for profiling genome-wide DNA methylation patterns in eukaryotes. RRBS couples size selection, bisulfite conversion, and second-generation sequencing to enrich for CpG-dense regions of the genome. The progressive improvement of second-generation sequencing technologies and reduction in cost provided an opportunity to examine the DNA methylation patterns of multiple genomes. Here, we describe a protocol for sequencing multiple RRBS libraries in a single sequencing reaction to generate base-resolution methylomes. Furthermore, we provide a brief guideline for base-calling and data analysis of multiplexed RRBS libraries. These strategies will be useful to perform large-scale, genome-wide DNA methylation analysis.

  20. Advanced Applications of Next-Generation Sequencing Technologies to Orchid Biology.

    Science.gov (United States)

    Yeh, Chuan-Ming; Liu, Zhong-Jian; Tsai, Wen-Chieh

    2017-09-08

    Next-generation sequencing technologies are revolutionizing biology by permitting, transcriptome sequencing, whole-genome sequencing and resequencing, and genome-wide single nucleotide polymorphism profiling. Orchid research has benefited from this breakthrough, and a few orchid genomes are now available; new biological questions can be approached and new breeding strategies can be designed. The first part of this review describes the unique features of orchid biology. The second part provides an overview of the current next-generation sequencing platforms, many of which are already used in plant laboratories. The third part summarizes the state of orchid transcriptome and genome sequencing and illustrates current achievements. The genetic sequences currently obtained will not only provide a broad scope for the study of orchid biology, but also serves as a starting point for uncovering the mystery of orchid evolution.

  1. Next-generation phylogeography: a targeted approach for multilocus sequencing of non-model organisms.

    Directory of Open Access Journals (Sweden)

    Jonathan B Puritz

    Full Text Available The field of phylogeography has long since realized the need and utility of incorporating nuclear DNA (nDNA sequences into analyses. However, the use of nDNA sequence data, at the population level, has been hindered by technical laboratory difficulty, sequencing costs, and problematic analytical methods dealing with genotypic sequence data, especially in non-model organisms. Here, we present a method utilizing the 454 GS-FLX Titanium pyrosequencing platform with the capacity to simultaneously sequence two species of sea star (Meridiastra calcar and Parvulastra exigua at five different nDNA loci across 16 different populations of 20 individuals each per species. We compare results from 3 populations with traditional Sanger sequencing based methods, and demonstrate that this next-generation sequencing platform is more time and cost effective and more sensitive to rare variants than Sanger based sequencing. A crucial advantage is that the high coverage of clonally amplified sequences simplifies haplotype determination, even in highly polymorphic species. This targeted next-generation approach can greatly increase the use of nDNA sequence loci in phylogeographic and population genetic studies by mitigating many of the time, cost, and analytical issues associated with highly polymorphic, diploid sequence markers.

  2. Next-Generation Phylogeography: A Targeted Approach for Multilocus Sequencing of Non-Model Organisms

    Science.gov (United States)

    Puritz, Jonathan B.; Addison, Jason A.; Toonen, Robert J.

    2012-01-01

    The field of phylogeography has long since realized the need and utility of incorporating nuclear DNA (nDNA) sequences into analyses. However, the use of nDNA sequence data, at the population level, has been hindered by technical laboratory difficulty, sequencing costs, and problematic analytical methods dealing with genotypic sequence data, especially in non-model organisms. Here, we present a method utilizing the 454 GS-FLX Titanium pyrosequencing platform with the capacity to simultaneously sequence two species of sea star (Meridiastra calcar and Parvulastra exigua) at five different nDNA loci across 16 different populations of 20 individuals each per species. We compare results from 3 populations with traditional Sanger sequencing based methods, and demonstrate that this next-generation sequencing platform is more time and cost effective and more sensitive to rare variants than Sanger based sequencing. A crucial advantage is that the high coverage of clonally amplified sequences simplifies haplotype determination, even in highly polymorphic species. This targeted next-generation approach can greatly increase the use of nDNA sequence loci in phylogeographic and population genetic studies by mitigating many of the time, cost, and analytical issues associated with highly polymorphic, diploid sequence markers. PMID:22470543

  3. Next generation sequencing on patients with LGMD and nonspecific myopathies: findings associated with ANO5 mutations

    OpenAIRE

    2015-01-01

    We studied 786 undiagnosed patients with LGMD or nonspecific myopathic features to investigate the role of ANO5 mutations in limb-girdle muscular dystrophies (LGMDs) and in nonspecific myopathies using the next generation sequencing (NGS) approach. In 160 LGMD patients, we first sequenced hotspot exons 5 and 20 and then sequenced the remaining part of the coding region. Another 626 patients, recruited using broader inclusion criteria, were directly analyzed by targeted NGS. By combining NGS a...

  4. Now and next-generation sequencing techniques: future of sequence analysis using cloud computing.

    Science.gov (United States)

    Thakur, Radhe Shyam; Bandopadhyay, Rajib; Chaudhary, Bratati; Chatterjee, Sourav

    2012-01-01

    Advances in the field of sequencing techniques have resulted in the greatly accelerated production of huge sequence datasets. This presents immediate challenges in database maintenance at datacenters. It provides additional computational challenges in data mining and sequence analysis. Together these represent a significant overburden on traditional stand-alone computer resources, and to reach effective conclusions quickly and efficiently, the virtualization of the resources and computation on a pay-as-you-go concept (together termed "cloud computing") has recently appeared. The collective resources of the datacenter, including both hardware and software, can be available publicly, being then termed a public cloud, the resources being provided in a virtual mode to the clients who pay according to the resources they employ. Examples of public companies providing these resources include Amazon, Google, and Joyent. The computational workload is shifted to the provider, which also implements required hardware and software upgrades over time. A virtual environment is created in the cloud corresponding to the computational and data storage needs of the user via the internet. The task is then performed, the results transmitted to the user, and the environment finally deleted after all tasks are completed. In this discussion, we focus on the basics of cloud computing, and go on to analyze the prerequisites and overall working of clouds. Finally, the applications of cloud computing in biological systems, particularly in comparative genomics, genome informatics, and SNP detection are discussed with reference to traditional workflows.

  5. Now And Next Generation Sequencing Techniques: Future of Sequence Analysis using Cloud Computing

    Directory of Open Access Journals (Sweden)

    Radhe Shyam Thakur

    2012-12-01

    Full Text Available Advancements in the field of sequencing techniques resulted in the huge sequenced data to be produced at a very faster rate. It is going cumbersome for the datacenter to maintain the databases. Data mining and sequence analysis approaches needs to analyze the databases several times to reach any efficient conclusion. To cope with such overburden on computer resources and to reach efficient and effective conclusions quickly, the virtualization of the resources and computation on pay as you go concept was introduced and termed as cloud computing. The datacenter’s hardware and software is collectively known as cloud which when available publicly is termed as public cloud. The datacenter’s resources are provided in a virtual mode to the clients via a service provider like Amazon, Google and Joyent which charges on pay as you go manner. The workload is shifted to the provider which is maintained by the required hardware and software upgradation. The service provider manages it by upgrading the requirements in the virtual mode. Basically a virtual environment is created according to the need of the user by taking permission from datacenter via internet, the task is performed and the environment is deleted after the task is over. In this discussion, we are focusing on the basics of cloud computing, the prerequisites and overall working of clouds. Furthermore, briefly the applications of cloud computing in biological systems, especially in comparative genomics, genome informatics and SNP detection with reference to traditional workflow are discussed.

  6. ISO 17025 validation of a next-generation sequencing assay for relationship testing

    DEFF Research Database (Denmark)

    Buchard, Anders; Kampmann, Marie-Louise; Poulsen, Lena

    2016-01-01

    The HID-Ion AmpliSeq™ Identity Panel is a next-generation sequencing assay with 90 autosomal and 34 Y-chromosome SNPs that are amplified in one PCR step and subsequently sequenced using the Ion Personal Genome Machine (Ion PGM™) System. This assay was validated for relationship testing in our ISO...

  7. Comparison of techniques for quantification of next-generation sequencing libraries

    DEFF Research Database (Denmark)

    Hussing, Christian; Kampmann, Marie-Louise; Mogensen, Helle Smidt;

    2015-01-01

    To ensure efficient sequencing, the DNA of next-generation sequencing (NGS) libraries must be quantified correctly. Therefore, an accurate, sensitive and stable method for DNA quantification is crucial. In this study, seven different methods for DNA quantification were compared to each other by q...

  8. Genomic relationships computed from either next- generation sequence or array SNP data

    NARCIS (Netherlands)

    Perez Enciso, M.

    2014-01-01

    The use of sequence data in genomic prediction models is a topic of high interest, given the decreasing prices of current next'-generation sequencing technologies (NGS) and the theoretical possibility of directly interrogating the genomes for all causal mutations. Here, we compare by simulation how

  9. From FASTQ to Function: In Silico Methods for Processing Next-Generation Sequencing Data.

    Science.gov (United States)

    Preston, Mark D; Stabler, Richard A

    2016-01-01

    This chapter presents a method to process C. difficile whole-genome next-generation sequencing data straight from the sequencer. Quality control processing and de novo assembly of these data enable downstream analyses such as gene annotation and in silico multi-locus strain-type identification.

  10. Molecular diagnostics of aleutian mink disease virus: applied use of next generation sequencing and phylogenetics

    DEFF Research Database (Denmark)

    Hagberg, Emma Elisabeth

    such as next generation sequencing cheaper and more easily available. Whole genome sequencing and advanced phylogenetic analyses have successfully been applied to describe the molecular evolution and transmission patterns for viruses such as Foot and Mouth Disease Virus (FMDV), Ebola, and avian influenza virus...

  11. Exploring the potential of next-generation sequencing in detection of respiratory Viruses

    NARCIS (Netherlands)

    S. Prachayangprecha (Slinporn); C.M.E. Schapendonk (Claudia); M.P.G. Koopmans D.V.M. (Marion); A.D.M.E. Osterhaus (Albert); A. Schürch (Anita); S.D. Pas (Suzan); A.A. Eijck (Annemiek); Y. Poovorawan (Yong); B.L. Haagmans (Bart); S.L. Smits (Saskia)

    2014-01-01

    textabstractEfficient detection of human respiratory viral pathogens is crucial in the management of patients with acute respiratory tract infection. Sequence-independent amplification of nucleic acids combined with next-generation sequencing technology and bioinformatics analyses is a promising str

  12. Stochastic modeling and generation of synthetic sequences of hourly global solar irradiation at Quetta, Pakistan

    Energy Technology Data Exchange (ETDEWEB)

    Kamal, Lalarukh [Balochistan Univ., Dept. of Mathematics, Quetta (Pakistan); Jafri, Yasmin Zahra [Balochistan Univ., Dept. of Statistics, Quetta (Pakistan)

    1999-07-01

    Using hourly global radiation data at Quetta, Pakistan for 10 yr, an Autoregressive Moving Average (ARMA) process is fitted. Markov Transition Matrices have also been developed. These models are used for generating synthetic sequences for hourly radiations in MJ/m{sup 2} and that the generated sequences are compared with the observed data. We found the MTM approach relatively better as a simulator compared to ARMA modeling. (Author)

  13. NGS Catalog: A Database of Next Generation Sequencing Studies in Humans

    OpenAIRE

    Xia, Junfeng; Wang, Qingguo; Jia, Peilin; Wang, Bing; Pao, William; Zhao, Zhongming

    2012-01-01

    Next generation sequencing (NGS) technologies have been rapidly applied in biomedical and biological research since its advent only a few years ago, and they are expected to advance at an unprecedented pace in the following years. To provide the research community with a comprehensive NGS resource, we have developed the database Next Generation Sequencing Catalog (NGS Catalog, http://bioinfo.mc.vanderbilt.edu/NGS/index.html), a continually updated database that collects, curates and manages a...

  14. Level traversal binary tree in sequence storage and its application MA Jing-shan, QIN Yu-ping%顺序存储二叉树的遍历及其应用研究

    Institute of Scientific and Technical Information of China (English)

    马靖善; 秦玉平

    2013-01-01

    The sequence storage binary tree is used to dispose that binary tree form hard upon full .This the-sis introduced binary tree structure of sequence storage and it's advantage, the traversal method, level traversal algorithm and recursive traversal algorithm of sequence binary tree , and some simple application on level travers-al algorithm.%  顺序存储二叉树非常适用于二叉树的树形接近于满二叉树时的处理。本文介绍了二叉树的顺序存储结构及其优点、二叉树的遍历方法、顺序存储二叉树的层次遍历和递归遍历算法,以及层次遍历算法的一些简单应用。

  15. Binary mask programmable hologram.

    Science.gov (United States)

    Tsang, P W M; Poon, T-C; Zhou, Changhe; Cheung, K W K

    2012-11-19

    We report, for the first time, the concept and generation of a novel Fresnel hologram called the digital binary mask programmable hologram (BMPH). A BMPH is comprised of a static, high resolution binary grating that is overlaid with a lower resolution binary mask. The reconstructed image of the BMPH can be programmed to approximate a target image (including both intensity and depth information) by configuring the pattern of the binary mask with a simple genetic algorithm (SGA). As the low resolution binary mask can be realized with less stringent display technology, our method enables the development of simple and economical holographic video display.

  16. Next-generation sequencing of human mitochondrial reference genomes uncovers high heteroplasmy frequency.

    Directory of Open Access Journals (Sweden)

    Maria Ximena Sosa

    Full Text Available We describe methods for rapid sequencing of the entire human mitochondrial genome (mtgenome, which involve long-range PCR for specific amplification of the mtgenome, pyrosequencing, quantitative mapping of sequence reads to identify sequence variants and heteroplasmy, as well as de novo sequence assembly. These methods have been used to study 40 publicly available HapMap samples of European (CEU and African (YRI ancestry to demonstrate a sequencing error rate <5.63×10(-4, nucleotide diversity of 1.6×10(-3 for CEU and 3.7×10(-3 for YRI, patterns of sequence variation consistent with earlier studies, but a higher rate of heteroplasmy varying between 10% and 50%. These results demonstrate that next-generation sequencing technologies allow interrogation of the mitochondrial genome in greater depth than previously possible which may be of value in biology and medicine.

  17. General simulation algorithm for autocorrelated binary processes

    Science.gov (United States)

    Serinaldi, Francesco; Lombardo, Federico

    2017-02-01

    The apparent ubiquity of binary random processes in physics and many other fields has attracted considerable attention from the modeling community. However, generation of binary sequences with prescribed autocorrelation is a challenging task owing to the discrete nature of the marginal distributions, which makes the application of classical spectral techniques problematic. We show that such methods can effectively be used if we focus on the parent continuous process of beta distributed transition probabilities rather than on the target binary process. This change of paradigm results in a simulation procedure effectively embedding a spectrum-based iterative amplitude-adjusted Fourier transform method devised for continuous processes. The proposed algorithm is fully general, requires minimal assumptions, and can easily simulate binary signals with power-law and exponentially decaying autocorrelation functions corresponding, for instance, to Hurst-Kolmogorov and Markov processes. An application to rainfall intermittency shows that the proposed algorithm can also simulate surrogate data preserving the empirical autocorrelation.

  18. Exploring the potential of next-generation sequencing in detection of respiratory viruses.

    Science.gov (United States)

    Prachayangprecha, Slinporn; Schapendonk, Claudia M E; Koopmans, Marion P; Osterhaus, Albert D M E; Schürch, Anita C; Pas, Suzan D; van der Eijk, Annemiek A; Poovorawan, Yong; Haagmans, Bart L; Smits, Saskia L

    2014-10-01

    Efficient detection of human respiratory viral pathogens is crucial in the management of patients with acute respiratory tract infection. Sequence-independent amplification of nucleic acids combined with next-generation sequencing technology and bioinformatics analyses is a promising strategy for identifying pathogens in clinical and public health settings. It allows the characterization of hundreds of different known pathogens simultaneously and of novel pathogens that elude conventional testing. However, major hurdles for its routine use exist, including cost, turnaround time, and especially sensitivity of the assay, as the detection limit is dependent on viral load, host genetic material, and sequencing depth. To obtain insights into these aspects, we analyzed nasopharyngeal aspirates from a cohort of 81 Thai children with respiratory disease for the presence of respiratory viruses using a sequence-independent next-generation sequencing approach and routinely used diagnostic real-time reverse transcriptase PCR (real-time RT-PCR) assays. With respect to the detection of rhinovirus and human metapneumovirus, the next-generation sequencing approach was at least as sensitive as diagnostic real-time RT-PCR in this small cohort, whereas for bocavirus and enterovirus, next-generation sequencing was less sensitive than real-time RT-PCR. The advantage of the sequencing approach over real-time RT-PCR was the immediate availability of virus-typing information. Considering the development of platforms capable of generating more output data at declining costs, next-generation sequencing remains of interest for future virus diagnosis in clinical and public health settings and certainly as an additional tool when screening results from real-time RT-PCR are negative.

  19. Five simple guidelines for establishing basic authenticity and reliability of newly generated fungal ITS sequences

    Directory of Open Access Journals (Sweden)

    R. Henrik Nilsson

    2012-09-01

    Full Text Available Molecular data form an important research tool in most branches of mycology. A non-trivial proportion of the public fungal DNA sequences are, however, compromised in terms of quality and reliability, contributing noise and bias to sequence-borne inferences such as phylogenetic analysis, diversity assessment, and barcoding. In this paper we discuss various aspects and pitfalls of sequence quality assessment. Based on our observations, we provide a set of guidelines to assist in manual quality management of newly generated, near-full-length (Sanger-derived fungal ITS sequences and to some extent also sequences of shorter read lengths, other genes or markers, and groups of organisms. The guidelines are intentionally non-technical and do not require substantial bioinformatics skills or significant computational power. Despite their simple nature, we feel they would have caught the vast majority of the severely compromised ITS sequences in the public corpus. Our guidelines are nevertheless not infallible, and common sense and intuition remain important elements in the pursuit of compromised sequence data. The guidelines focus on basic sequence authenticity and reliability of the newly generated sequences, and the user may want to consider additional resources and steps to accomplish the best possible quality control. A discussion on the technical resources for further sequence quality management is therefore provided in the supplementary material.

  20. Monte Carlo simulations of post-common-envelope white dwarf + main sequence binaries: The effects of including recombination energy

    CERN Document Server

    Zorotovic, M; García-Berro, E; Camacho, J; Torres, S; Rebassa-Mansergas, A; Gänsicke, B T

    2014-01-01

    Detached WD+MS PCEBs are perhaps the most suitable objects for testing predictions of close-compact binary-star evolution theories, in particular, CE evolution. The population of WD+MS PCEBs has been simulated by several authors in the past and compared with observations. However, most of those predictions did not take the possible contributions to the envelope ejection from additional sources of energy (mostly recombination energy) into account. Here we update existing binary population models of WD+MS PCEBs by assuming that a fraction of the recombination energy available within the envelope contributes to ejecting the envelope. We performed Monte Carlo simulations of 10^7 MS+MS binaries for 9 different models using standard assumptions for the initial primary mass function, binary separations, and initial-mass-ratio distribution and evolved these systems using the publicly available BSE code. Including a fraction of recombination energy leads to a clear prediction of a large number of long orbital period (...

  1. Processing Binary and Fuzzy Logic by Chaotic Time Series Generated by a Hydrodynamic Photochemical Oscillator.

    Science.gov (United States)

    Gentili, Pier Luigi; Giubila, Maria Sole; Heron, B Mark

    2017-02-03

    This work demonstrates the computational power of a hydrodynamic photochemical oscillator based on a photochromic naphthopyran, generating aperiodic time series. The chaotic character of the time series is tested by calculating its largest Lyapunov exponent and the correlation dimension of its attractor after building its phase space through the Takens' theorem. Then, the chaotic dynamic is shown to be suitable to implement all the fundamental Boolean two-inputs-one-output logic gates. Finally, the strategy to implement Fuzzy logic systems (FLSs) based on the time series is described. Such FLSs promise to be useful in the field of Computational Linguistics that is concerned with the development of artificial intelligent systems able to transform collections of numerical data in natural language texts.

  2. Next generation sequencing in clinical medicine: Challenges and lessons for pathology and biomedical informatics

    Directory of Open Access Journals (Sweden)

    Rama R Gullapalli

    2012-01-01

    Full Text Available The Human Genome Project (HGP provided the initial draft of mankind′s DNA sequence in 2001. The HGP was produced by 23 collaborating laboratories using Sanger sequencing of mapped regions as well as shotgun sequencing techniques in a process that occupied 13 years at a cost of ~$3 billion. Today, Next Generation Sequencing (NGS techniques represent the next phase in the evolution of DNA sequencing technology at dramatically reduced cost compared to traditional Sanger sequencing. A single laboratory today can sequence the entire human genome in a few days for a few thousand dollars in reagents and staff time. Routine whole exome or even whole genome sequencing of clinical patients is well within the realm of affordability for many academic institutions across the country. This paper reviews current sequencing technology methods and upcoming advancements in sequencing technology as well as challenges associated with data generation, data manipulation and data storage. Implementation of routine NGS data in cancer genomics is discussed along with potential pitfalls in the interpretation of the NGS data. The overarching importance of bioinformatics in the clinical implementation of NGS is emphasized. [7] We also review the issue of physician education which also is an important consideration for the successful implementation of NGS in the clinical workplace. NGS technologies represent a golden opportunity for the next generation of pathologists to be at the leading edge of the personalized medicine approaches coming our way. Often under-emphasized issues of data access and control as well as potential ethical implications of whole genome NGS sequencing are also discussed. Despite some challenges, it′s hard not to be optimistic about the future of personalized genome sequencing and its potential impact on patient care and the advancement of knowledge of human biology and disease in the near future.

  3. Monte Carlo simulations of post-common-envelope white dwarf + main sequence binaries: comparison with the SDSS DR7 observed sample

    CERN Document Server

    Camacho, J; García-Berro, E; Zorotovic, M; Schreiber, M R; Rebassa-Mansergas, A; Gómez-Morán, A Nebot; Gänsicke, B T

    2014-01-01

    Detached white dwarf + main sequence (WD+MS) systems represent the simplest population of post-common envelope binaries (PCEBs). Since the ensemble properties of this population carries important information about the characteristics of the common-envelope (CE) phase, it deserves close scrutiny. However, most population synthesis studies do not fully take into account the effects of the observational selection biases of the samples used to compare with the theoretical simulations. Here we present the results of a set of detailed Monte Carlo simulations of the population of WD+MS binaries in the Sloan Digital Sky Survey (SDSS) Data Release 7. We used up-to-date stellar evolutionary models, a complete treatment of the Roche lobe overflow episode, and a full implementation of the orbital evolution of the binary systems. Moreover, in our treatment we took into account the selection criteria and all the known observational biases. Our population synthesis study allowed us to make a meaningful comparison with the a...

  4. Genomic and Functional Characteristics of Human Cytomegalovirus Revealed by Next-Generation Sequencing

    Directory of Open Access Journals (Sweden)

    Steven Sijmons

    2014-03-01

    Full Text Available The complete genome of human cytomegalovirus (HCMV was elucidated almost 25 years ago using a traditional cloning and Sanger sequencing approach. Analysis of the genetic content of additional laboratory and clinical isolates has lead to a better, albeit still incomplete, definition of the coding potential and diversity of wild-type HCMV strains. The introduction of a new generation of massively parallel sequencing technologies, collectively called next-generation sequencing, has profoundly increased the throughput and resolution of the genomics field. These increased possibilities are already leading to a better understanding of the circulating diversity of HCMV clinical isolates. The higher resolution of next-generation sequencing provides new opportunities in the study of intrahost viral population structures. Furthermore, deep sequencing enables novel diagnostic applications for sensitive drug resistance mutation detection. RNA-seq applications have changed the picture of the HCMV transcriptome, which resulted in proof of a vast amount of splicing events and alternative transcripts. This review discusses the application of next-generation sequencing technologies, which has provided a clearer picture of the intricate nature of the HCMV genome. The continuing development and application of novel sequencing technologies will further augment our understanding of this ubiquitous, but elusive, herpesvirus.

  5. Sequence Alignment with Dynamic Divisor Generation for Keystroke Dynamics Based User Authentication

    Directory of Open Access Journals (Sweden)

    Jiacang Ho

    2015-01-01

    Full Text Available Keystroke dynamics based authentication is one of the prevention mechanisms used to protect one’s account from criminals’ illegal access. In this authentication mechanism, keystroke dynamics are used to capture patterns in a user typing behavior. Sequence alignment is shown to be one of effective algorithms for keystroke dynamics based authentication, by comparing the sequences of keystroke data to detect imposter’s anomalous sequences. In previous research, static divisor has been used for sequence generation from the keystroke data, which is a number used to divide a time difference of keystroke data into an equal-length subinterval. After the division, the subintervals are mapped to alphabet letters to form sequences. One major drawback of this static divisor is that the amount of data for this subinterval generation is often insufficient, which leads to premature termination of subinterval generation and consequently causes inaccurate sequence alignment. To alleviate this problem, we introduce sequence alignment of dynamic divisor (SADD in this paper. In SADD, we use mean of Horner’s rule technique to generate dynamic divisors and apply them to produce the subintervals with different length. The comparative experimental results with SADD and other existing algorithms indicate that SADD is usually comparable to and often outperforms other existing algorithms.

  6. Historical perspective, development and applications of next-generation sequencing in plant virology.

    Science.gov (United States)

    Barba, Marina; Czosnek, Henryk; Hadidi, Ahmed

    2014-01-06

    Next-generation high throughput sequencing technologies became available at the onset of the 21st century. They provide a highly efficient, rapid, and low cost DNA sequencing platform beyond the reach of the standard and traditional DNA sequencing technologies developed in the late 1970s. They are continually improved to become faster, more efficient and cheaper. They have been used in many fields of biology since 2004. In 2009, next-generation sequencing (NGS) technologies began to be applied to several areas of plant virology including virus/viroid genome sequencing, discovery and detection, ecology and epidemiology, replication and transcription. Identification and characterization of known and unknown viruses and/or viroids in infected plants are currently among the most successful applications of these technologies. It is expected that NGS will play very significant roles in many research and non-research areas of plant virology.

  7. Applications and case studies of the next-generation sequencing technologies in food, nutrition and agriculture.

    Science.gov (United States)

    Liu, George E

    2009-01-01

    The next-generation sequencing technologies are able to produce millions of short sequence reads in a high-throughput, cost-effective fashion. The emergence of these technologies has not only facilitated genome sequencing but also started to change the landscape of life sciences. Here, I survey their major applications ranging from whole-genome sequencing and resequencing, single nucleotide polymorphism (SNP) and structural variation discovery, to mRNA and noncoding RNA profiling and protein-nucleic acid interaction assay. These case studies in structural, functional and comparative genomics, metagenomics, and epigenomics are providing a more complete picture of the genome structures and functions. In the near future, we will witness broad impacts of these next-generation sequencing technologies for solving the complex biological problems in food, nutrition and agriculture. In this article, recent patents based information is also included.

  8. Historical Perspective, Development and Applications of Next-Generation Sequencing in Plant Virology

    Directory of Open Access Journals (Sweden)

    Marina Barba

    2014-01-01

    Full Text Available Next-generation high throughput sequencing technologies became available at the onset of the 21st century. They provide a highly efficient, rapid, and low cost DNA sequencing platform beyond the reach of the standard and traditional DNA sequencing technologies developed in the late 1970s. They are continually improved to become faster, more efficient and cheaper. They have been used in many fields of biology since 2004. In 2009, next-generation sequencing (NGS technologies began to be applied to several areas of plant virology including virus/viroid genome sequencing, discovery and detection, ecology and epidemiology, replication and transcription. Identification and characterization of known and unknown viruses and/or viroids in infected plants are currently among the most successful applications of these technologies. It is expected that NGS will play very significant roles in many research and non-research areas of plant virology.

  9. Terminal sequence importance of de novo proteins from binary-patterned library: stable artificial proteins with 11- or 12-amino acid alphabet.

    Science.gov (United States)

    Okura, Hiromichi; Takahashi, Tsuyoshi; Mihara, Hisakazu

    2012-06-01

    Successful approaches of de novo protein design suggest a great potential to create novel structural folds and to understand natural rules of protein folding. For these purposes, smaller and simpler de novo proteins have been developed. Here, we constructed smaller proteins by removing the terminal sequences from stable de novo vTAJ proteins and compared stabilities between mutant and original proteins. vTAJ proteins were screened from an α3β3 binary-patterned library which was designed with polar/ nonpolar periodicities of α-helix and β-sheet. vTAJ proteins have the additional terminal sequences due to the method of constructing the genetically repeated library sequences. By removing the parts of the sequences, we successfully obtained the stable smaller de novo protein mutants with fewer amino acid alphabets than the originals. However, these mutants showed the differences on ANS binding properties and stabilities against denaturant and pH change. The terminal sequences, which were designed just as flexible linkers not as secondary structure units, sufficiently affected these physicochemical details. This study showed implications for adjusting protein stabilities by designing N- and C-terminal sequences.

  10. Structural searches using isopointal sets as generators: densest packings for binary hard sphere mixtures.

    Science.gov (United States)

    Hudson, Toby S; Harrowell, Peter

    2011-05-18

    Algorithms to search for crystal structures that optimize some extensive property (energy, volume, etc) typically make use of random particle reorganizations in the context of one or more numerical techniques such as simulated annealing, genetic algorithms or biased random walks, applied to the coordinates of every particle in the unit cell, together with the cell angles and lengths. In this paper we describe the restriction of such searches to predefined isopointal sets, breaking the problem into countable sub-problems which exploit crystal symmetries to reduce the dimensionality of the search space. Applying this method to the search for maximally packed mixtures of hard spheres of two sizes, we demonstrate that the densest packed structures can be identified by searches within a couple of isopointal sets. For the A(2)B system, the densest known packings over the entire tested range 0.2 < r(A)/r(B) < 2.5, including some improvements on previous optima, can all be identified by searches within a single isopointal set. In the case of the AB composition, searches of two isopointal sets generate the densest packed structures over the radius ratio range 0.2 < r(A)/r(B) < 5.0.

  11. Evaluation ofA Single-reaction Method for Whole Genome Sequencing of Influenza A Virus using Next Generation Sequencing

    Institute of Scientific and Technical Information of China (English)

    ZOU Xiao Hui; CHEN Wen Bing; ZHAO Xiang; ZHU Wen Fei; YANG Lei; WANG Da Yan; SHU Yue Long

    2016-01-01

    ObjectiveTo evaluate a single-reaction genome amplification method, the multisegment reverse transcription-PCR (M-RTPCR), for its sensitivity to full genome sequencing of influenza A virus, and the ability to differentiate mix-subtype virus, using the next generation sequencing (NGS) platform. MethodsVirus genome copy was quantified and serially diluted to different titers, followed by amplification with the M-RTPCR method and sequencing on the NGS platform. Furthermore, we manually mixed two subtype viruses to different titer rate and amplified the mixed virus with the M-RTPCR protocol, followed by whole genome sequencing on the NGS platform. We also used clinical samples to test the method performance. ResultsThe M-RTPCR method obtained complete genome of testing virus at 125 copies/reaction and determined the virus subtype at titer of 25 copies/reaction. Moreover, the two subtypes in the mixed virus could be discriminated, even though these two virus copies differed by 200-fold using this amplification protocol. The sensitivity of this protocol we detected using virus RNA was also confirmed with clinical samples containing low-titer virus. ConclusionThe M-RTPCR is a robust and sensitive amplification method for whole genome sequencing of influenza A virus using NGS platform.

  12. Next-Generation Sequencing Workflow for NSCLC Critical Samples Using a Targeted Sequencing Approach by Ion Torrent PGM™ Platform

    Directory of Open Access Journals (Sweden)

    Irene Vanni

    2015-12-01

    Full Text Available Next-generation sequencing (NGS is a cost-effective technology capable of screening several genes simultaneously; however, its application in a clinical context requires an established workflow to acquire reliable sequencing results. Here, we report an optimized NGS workflow analyzing 22 lung cancer-related genes to sequence critical samples such as DNA from formalin-fixed paraffin-embedded (FFPE blocks and circulating free DNA (cfDNA. Snap frozen and matched FFPE gDNA from 12 non-small cell lung cancer (NSCLC patients, whose gDNA fragmentation status was previously evaluated using a multiplex PCR-based quality control, were successfully sequenced with Ion Torrent PGM™. The robust bioinformatic pipeline allowed us to correctly call both Single Nucleotide Variants (SNVs and indels with a detection limit of 5%, achieving 100% specificity and 96% sensitivity. This workflow was also validated in 13 FFPE NSCLC biopsies. Furthermore, a specific protocol for low input gDNA capable of producing good sequencing data with high coverage, high uniformity, and a low error rate was also optimized. In conclusion, we demonstrate the feasibility of obtaining gDNA from FFPE samples suitable for NGS by performing appropriate quality controls. The optimized workflow, capable of screening low input gDNA, highlights NGS as a potential tool in the detection, disease monitoring, and treatment of NSCLC.

  13. HAPCAD: An open-source tool to detect PCR crossovers in next-generation sequencing generated HLA data.

    Science.gov (United States)

    McDevitt, Shana L; Bredeson, Jessen V; Roy, Scott W; Lane, Julie A; Noble, Janelle A

    2016-03-01

    Next-generation sequencing (NGS) based HLA genotyping can generate PCR artifacts corresponding to IMGT/HLA Database alleles, for which multiple examples have been observed, including sequence corresponding to the HLA-DRB1(∗)03:42 allele. Repeat genotyping of 131 samples, previously genotyped as DRB1(∗)03:01 homozygotes using probe-based methods, resulted in the heterozygous call DRB1(∗)03:01+DRB1(∗)03:42. The apparent rare DRB1(∗)03:42 allele is hypothesized to be a "hybrid amplicon" generated by PCR crossover, a process in which a partial PCR product denatures from its template, anneals to a different allele template, and extends to completion. Unlike most PCR crossover products, "hybrid amplicons" always corresponds to an IMGT/HLA Database allele, necessitating a case-by-case analysis of whether its occurrence reflects the actual allele or is simply the result of PCR crossover. The Hybrid Amplicon/PCR Crossover Artifact Detector (HAPCAD) program mimics jumping PCR in silico and flags allele sequences that may also be generated as hybrid amplicon.

  14. Optimization of binary sequences based on evolutionary algorithm%基于进化计算的二元序列优化算法研究

    Institute of Scientific and Technical Information of China (English)

    李鹤; 李琦; 高军萍; 雷明然

    2014-01-01

    具有良好非周期自相关特性二元序列在通信同步、雷达等领域具有广泛的应用。通过对遗传算法、粒子群算法与量子粒子群算法三种进化算法进行对比分析,设计了具有良好非周期自相关特性的二元序列的搜索算法。研究结果表明,粒子群算法的搜索能力优于遗传算法,而量子粒子群算法具有参数少,易于控制的优点,取得了较好的优化结果。%Binary sequences with good aperiodic autocorrelation features are widely used in the field of radar, communication synchronization. Genetic algorithm, particle swarm optimization and quantum particle swarm optimization algorithm are compared and analyzed in this paper. The new search algorithm of binary sequences with good aperiodic autocorrelation properties are designed based on three evolutionary algorithms. Research results show that the search ability of particle swarm algorithm is better than genetic algorithm. Quantum particle swarm optimization algorithm has less parameters, easy to control, and the better good optimization results were obtained.

  15. Genomic sequencing identifies novel Bacillus thuringiensis Vip1/Vip2 binary and Cry8 toxins that have high toxicity to Scarabaeoidea larvae.

    Science.gov (United States)

    Bi, Yang; Zhang, Yanrui; Shu, Changlong; Crickmore, Neil; Wang, Qinglei; Du, Lixin; Song, Fuping; Zhang, Jie

    2015-01-01

    The Bacillus thuringiensis strain HBF-18 (CGMCC 2070), which has previously been shown to encode the cry8Ga toxin gene, is active against both Holotrichia oblita and Holotrichia parallela. Recombinant Cry8Ga however is only weakly toxic to these insect pests suggesting the involvement of additional toxins in the native strain. We report that through the use of Illumina sequencing three additional, and novel, genes, namely vip1Ad1, vip2Ag1, and cry8-like, were identified in this strain. Although no protein corresponding to these genes could be identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the HBF-18 proteome, reverse transcription (RT)-PCR indicated that all three genes were transcribed in the native strain. The two vip genes were cloned and expressed and, as with other Vip1/2 toxins, appeared to function as a binary toxin and showed strong activity against H. oblita, H. parallela and Anomala corpulenta. This is the first report to demonstrate that the Vip1/Vip2 binary toxin is active against these Scarabaeoidea larvae. The cry8-like gene appeared to be a C-terminally truncated form of a typical cry8 gene and was not expressed in our usual recombinant Bt expression system. When however the missing C-terminal region was replaced with the corresponding sequence from cry8Ea, the resulting hybrid expressed well and the toxin was active against the three test insects.

  16. Model-based quality assessment and base-calling for second-generation sequencing data.

    Science.gov (United States)

    Bravo, Héctor Corrada; Irizarry, Rafael A

    2010-09-01

    Second-generation sequencing (sec-gen) technology can sequence millions of short fragments of DNA in parallel, making it capable of assembling complex genomes for a small fraction of the price and time of previous technologies. In fact, a recently formed international consortium, the 1000 Genomes Project, plans to fully sequence the genomes of approximately 1200 people. The prospect of comparative analysis at the sequence level of a large number of samples across multiple populations may be achieved within the next five years. These data present unprecedented challenges in statistical analysis. For instance, analysis operates on millions of short nucleotide sequences, or reads-strings of A,C,G, or T's, between 30 and 100 characters long-which are the result of complex processing of noisy continuous fluorescence intensity measurements known as base-calling. The complexity of the base-calling discretization process results in reads of widely varying quality within and across sequence samples. This variation in processing quality results in infrequent but systematic errors that we have found to mislead downstream analysis of the discretized sequence read data. For instance, a central goal of the 1000 Genomes Project is to quantify across-sample variation at the single nucleotide level. At this resolution, small error rates in sequencing prove significant, especially for rare variants. Sec-gen sequencing is a relatively new technology for which potential biases and sources of obscuring variation are not yet fully understood. Therefore, modeling and quantifying the uncertainty inherent in the generation of sequence reads is of utmost importance. In this article, we present a simple model to capture uncertainty arising in the base-calling procedure of the Illumina/Solexa GA platform. Model parameters have a straightforward interpretation in terms of the chemistry of base-calling allowing for informative and easily interpretable metrics that capture the variability in

  17. Quick genetic screening using targeted next-generation sequencing in patients with tuberous sclerosis.

    Science.gov (United States)

    Liu, Qing; Huang, Yan; Zhang, Mingrong; Wang, Lian Qing; Guo, Xia Nan; Si, Nuo; Qi, Zhan; Zhou, Xiang Qin; Cui, Li-ying

    2015-04-01

    Tuberous sclerosis complex is an autosomal dominant disorder characterized by hamartomas in multiple organ systems. Mutations in the 2 large genes TSC1 and TSC2 have been demonstrated to be associated with tuberous sclerosis complex by various mutation screening methods. Targeted next-generation sequencing for genetic analysis is performed in the current study and is proved to be less cost, labor, and time consuming compared with Sanger sequencing. Two de novo and 1 recurrent TSC2 mutation in patients with tuberous sclerosis complex were revealed. Clinical details of patients were described and the underlying mechanism of the 2 novel TSC2 mutations, c.245G>A(p.W82X) and c.5405_5408dupACTT(p.P1803Lfs*25), were discussed. These results added to variability of TSC mutation spectrum and suggest that targeted next-generation sequencing could be the primary choice over Sanger sequencing in future tuberous sclerosis complex genetic counseling.

  18. NeSSM: a Next-generation Sequencing Simulator for Metagenomics.

    Directory of Open Access Journals (Sweden)

    Ben Jia

    Full Text Available BACKGROUND: Metagenomics can reveal the vast majority of microbes that have been missed by traditional cultivation-based methods. Due to its extremely wide range of application areas, fast metagenome sequencing simulation systems with high fidelity are in great demand to facilitate the development and comparison of metagenomics analysis tools. RESULTS: We present here a customizable metagenome simulation system: NeSSM (Next-generation Sequencing Simulator for Metagenomics. Combining complete genomes currently available, a community composition table, and sequencing parameters, it can simulate metagenome sequencing better than existing systems. Sequencing error models based on the explicit distribution of errors at each base and sequencing coverage bias are incorporated in the simulation. In order to improve the fidelity of simulation, tools are provided by NeSSM to estimate the sequencing error models, sequencing coverage bias and the community composition directly from existing metagenome sequencing data. Currently, NeSSM supports single-end and pair-end sequencing for both 454 and Illumina platforms. In addition, a GPU (graphics processing units version of NeSSM is also developed to accelerate the simulation. By comparing the simulated sequencing data from NeSSM with experimental metagenome sequencing data, we have demonstrated that NeSSM performs better in many aspects than existing popular metagenome simulators, such as MetaSim, GemSIM and Grinder. The GPU version of NeSSM is more than one-order of magnitude faster than MetaSim. CONCLUSIONS: NeSSM is a fast simulation system for high-throughput metagenome sequencing. It can be helpful to develop tools and evaluate strategies for metagenomics analysis and it's freely available for academic users at http://cbb.sjtu.edu.cn/~ccwei/pub/software/NeSSM.php.

  19. Z-DNA-forming sequences generate large-scale deletions in mammalian cells

    OpenAIRE

    Wang, Guliang; Christensen, Laura A.; Vasquez, Karen M.

    2006-01-01

    Spontaneous chromosomal breakages frequently occur at genomic hot spots in the absence of DNA damage and can result in translocation-related human disease. Chromosomal breakpoints are often mapped near purine–pyrimidine Z-DNA-forming sequences in human tumors. However, it is not known whether Z-DNA plays a role in the generation of these chromosomal breakages. Here, we show that Z-DNA-forming sequences induce high levels of genetic instability in both bacterial and mammalian cells. In mammali...

  20. An integrated SNP mining and utilization (ISMU pipeline for next generation sequencing data.

    Directory of Open Access Journals (Sweden)

    Sarwar Azam

    Full Text Available Open source single nucleotide polymorphism (SNP discovery pipelines for next generation sequencing data commonly requires working knowledge of command line interface, massive computational resources and expertise which is a daunting task for biologists. Further, the SNP information generated may not be readily used for downstream processes such as genotyping. Hence, a comprehensive pipeline has been developed by integrating several open source next generation sequencing (NGS tools along with a graphical user interface called Integrated SNP Mining and Utilization (ISMU for SNP discovery and their utilization by developing genotyping assays. The pipeline features functionalities such as pre-processing of raw data, integration of open source alignment tools (Bowtie2, BWA, Maq, NovoAlign and SOAP2, SNP prediction (SAMtools/SOAPsnp/CNS2snp and CbCC methods and interfaces for developing genotyping assays. The pipeline outputs a list of high quality SNPs between all pairwise combinations of genotypes analyzed, in addition to the reference genome/sequence. Visualization tools (Tablet and Flapjack integrated into the pipeline enable inspection of the alignment and errors, if any. The pipeline also provides a confidence score or polymorphism information content value with flanking sequences for identified SNPs in standard format required for developing marker genotyping (KASP and Golden Gate assays. The pipeline enables users to process a range of NGS datasets such as whole genome re-sequencing, restriction site associated DNA sequencing and transcriptome sequencing data at a fast speed. The pipeline is very useful for plant genetics and breeding community with no computational expertise in order to discover SNPs and utilize in genomics, genetics and breeding studies. The pipeline has been parallelized to process huge datasets of next generation sequencing. It has been developed in Java language and is available at http://hpc.icrisat.cgiar.org/ISMU as a

  1. Transcriptome analysis of carnation (Dianthus caryophyllus L. based on next-generation sequencing technology

    Directory of Open Access Journals (Sweden)

    Tanase Koji

    2012-07-01

    Full Text Available Abstract Background Carnation (Dianthus caryophyllus L., in the family Caryophyllaceae, can be found in a wide range of colors and is a model system for studies of flower senescence. In addition, it is one of the most important flowers in the global floriculture industry. However, few genomics resources, such as sequences and markers are available for carnation or other members of the Caryophyllaceae. To increase our understanding of the genetic control of important characters in carnation, we generated an expressed sequence tag (EST database for a carnation cultivar important in horticulture by high-throughput sequencing using 454 pyrosequencing technology. Results We constructed a normalized cDNA library and a 3’-UTR library of carnation, obtaining a total of 1,162,126 high-quality reads. These reads were assembled into 300,740 unigenes consisting of 37,844 contigs and 262,896 singlets. The contigs were searched against an Arabidopsis sequence database, and 61.8% (23,380 of them had at least one BLASTX hit. These contigs were also annotated with Gene Ontology (GO and were found to cover a broad range of GO categories. Furthermore, we identified 17,362 potential simple sequence repeats (SSRs in 14,291 of the unigenes. We focused on gene discovery in the areas of flower color and ethylene biosynthesis. Transcripts were identified for almost every gene involved in flower chlorophyll and carotenoid metabolism and in anthocyanin biosynthesis. Transcripts were also identified for every step in the ethylene biosynthesis pathway. Conclusions We present the first large-scale sequence data set for carnation, generated using next-generation sequencing technology. The large EST database generated from these sequences is an informative resource for identifying genes involved in various biological processes in carnation and provides an EST resource for understanding the genetic diversity of this plant.

  2. Improved Detection by Next-Generation Sequencing of Pyrazinamide Resistance in Mycobacterium tuberculosis Isolates

    OpenAIRE

    Maningi, Nontuthuko E.; Daum, Luke T.; Rodriguez, John D.; Mphahlele, Matsie; Peters, Remco P. H.; Fischer, Gerald W.; Chambers, James P.; Fourie, P. Bernard

    2015-01-01

    The technical limitations of common tests used for detecting pyrazinamide (PZA) resistance in Mycobacterium tuberculosis isolates pose challenges for comprehensive and accurate descriptions of drug resistance in patients with multidrug-resistant tuberculosis (MDR-TB). In this study, a 606-bp fragment (comprising the pncA coding region plus the promoter) was sequenced using Ion Torrent next-generation sequencing (NGS) to detect associated PZA resistance mutations in 88 recultured MDR-TB isolat...

  3. Low Diversity in the Mitogenome of Sperm Whales Revealed by Next-Generation Sequencing

    OpenAIRE

    Alexander, Alana; Steel, Debbie; Slikas, Beth; Hoekzema, Kendra; Carraher, Colm; Parks, Matthew; Cronn, Richard; Baker, C. Scott

    2012-01-01

    Large population sizes and global distributions generally associate with high mitochondrial DNA control region (CR) diversity. The sperm whale (Physeter macrocephalus) is an exception, showing low CR diversity relative to other cetaceans; however, diversity levels throughout the remainder of the sperm whale mitogenome are unknown. We sequenced 20 mitogenomes from 17 sperm whales representative of worldwide diversity using Next Generation Sequencing (NGS) technologies (Illumina GAIIx, Roche 45...

  4. Internally generated sequences in learning and executing goal-directed behavior.

    Science.gov (United States)

    Pezzulo, Giovanni; van der Meer, Matthijs A A; Lansink, Carien S; Pennartz, Cyriel M A

    2014-12-01

    A network of brain structures including hippocampus (HC), prefrontal cortex, and striatum controls goal-directed behavior and decision making. However, the neural mechanisms underlying these functions are unknown. Here, we review the role of 'internally generated sequences': structured, multi-neuron firing patterns in the network that are not confined to signaling the current state or location of an agent, but are generated on the basis of internal brain dynamics. Neurophysiological studies suggest that such sequences fulfill functions in memory consolidation, augmentation of representations, internal simulation, and recombination of acquired information. Using computational modeling, we propose that internally generated sequences may be productively considered a component of goal-directed decision systems, implementing a sampling-based inference engine that optimizes goal acquisition at multiple timescales of on-line choice, action control, and learning.

  5. [Next generation sequencing and its applications in non-invasive prenatal testing of aneuploidies].

    Science.gov (United States)

    Babay, Lilla Éva; Horányi, Dániel; Rigó, János; Nagy, Gyula Richárd

    2015-06-28

    The development of the new generation sequencing techniques brought a new era in the field of DNA sequencing, that also revolutionized the prenatal screening for aneuploidy. In order to provide a more complete view, the authors describe some first generation methods as well as the theoretical and technical background of the next generation methods. In the second part of this review, the authors focuse on non-invasive prenatal testing, which is a fetal cell-free DNA based method requiring advanced sequencing procedures. After discussing the theoretical and technical background, the authors review current application and utility of non-invasive prenatal testing. They conclude that non-invasive prenatal testing is the most effective screening test in high risk pregnancies and its efficiency can be justified in studies involving low risk pregnancies as well.

  6. FLEXBAR—Flexible Barcode and Adapter Processing for Next-Generation Sequencing Platforms

    Directory of Open Access Journals (Sweden)

    Matthias Dodt

    2012-12-01

    Full Text Available Quantitative and systems biology approaches benefit from the unprecedented depth of next-generation sequencing. A typical experiment yields millions of short reads, which oftentimes carry particular sequence tags. These tags may be: (a specific to the sequencing platform and library construction method (e.g., adapter sequences; (b have been introduced by experimental design (e.g., sample barcodes; or (c constitute some biological signal (e.g., splice leader sequences in nematodes. Our software FLEXBAR enables accurate recognition, sorting and trimming of sequence tags with maximal flexibility, based on exact overlap sequence alignment. The software supports data formats from all current sequencing platforms, including color-space reads. FLEXBAR maintains read pairings and processes separate barcode reads on demand. Our software facilitates the fine-grained adjustment of sequence tag detection parameters and search regions. FLEXBAR is a multi-threaded software and combines speed with precision. Even complex read processing scenarios might be executed with a single command line call. We demonstrate the utility of the software in terms of read mapping applications, library demultiplexing and splice leader detection. FLEXBAR and additional information is available for academic use from the website: http://sourceforge.net/projects/flexbar/.

  7. Molecular characterization of transgene integration by next-generation sequencing in transgenic cattle.

    Directory of Open Access Journals (Sweden)

    Ran Zhang

    Full Text Available As the number of transgenic livestock increases, reliable detection and molecular characterization of transgene integration sites and copy number are crucial not only for interpreting the relationship between the integration site and the specific phenotype but also for commercial and economic demands. However, the ability of conventional PCR techniques to detect incomplete and multiple integration events is limited, making it technically challenging to characterize transgenes. Next-generation sequencing has enabled cost-effective, routine and widespread high-throughput genomic analysis. Here, we demonstrate the use of next-generation sequencing to extensively characterize cattle harboring a 150-kb human lactoferrin transgene that was initially analyzed by chromosome walking without success. Using this approach, the sites upstream and downstream of the target gene integration site in the host genome were identified at the single nucleotide level. The sequencing result was verified by event-specific PCR for the integration sites and FISH for the chromosomal location. Sequencing depth analysis revealed that multiple copies of the incomplete target gene and the vector backbone were present in the host genome. Upon integration, complex recombination was also observed between the target gene and the vector backbone. These findings indicate that next-generation sequencing is a reliable and accurate approach for the molecular characterization of the transgene sequence, integration sites and copy number in transgenic species.

  8. Generation of novel motor sequences: the neural correlates of musical improvisation.

    Science.gov (United States)

    Berkowitz, Aaron L; Ansari, Daniel

    2008-06-01

    While some motor behavior is instinctive and stereotyped or learned and re-executed, much action is a spontaneous response to a novel set of environmental conditions. The neural correlates of both pre-learned and cued motor sequences have been previously studied, but novel motor behavior has thus far not been examined through brain imaging. In this paper, we report a study of musical improvisation in trained pianists with functional magnetic resonance imaging (fMRI), using improvisation as a case study of novel action generation. We demonstrate that both rhythmic (temporal) and melodic (ordinal) motor sequence creation modulate activity in a network of brain regions comprised of the dorsal premotor cortex, the rostral cingulate zone of the anterior cingulate cortex, and the inferior frontal gyrus. These findings are consistent with a role for the dorsal premotor cortex in movement coordination, the rostral cingulate zone in voluntary selection, and the inferior frontal gyrus in sequence generation. Thus, the invention of novel motor sequences in musical improvisation recruits a network of brain regions coordinated to generate possible sequences, select among them, and execute the decided-upon sequence.

  9. Analysis of quality raw data of second generation sequencers with Quality Assessment Software

    Directory of Open Access Journals (Sweden)

    Schneider Maria PC

    2011-04-01

    Full Text Available Abstract Background Second generation technologies have advantages over Sanger; however, they have resulted in new challenges for the genome construction process, especially because of the small size of the reads, despite the high degree of coverage. Independent of the program chosen for the construction process, DNA sequences are superimposed, based on identity, to extend the reads, generating contigs; mismatches indicate a lack of homology and are not included. This process improves our confidence in the sequences that are generated. Findings We developed Quality Assessment Software, with which one can review graphs showing the distribution of quality values from the sequencing reads. This software allow us to adopt more stringent quality standards for sequence data, based on quality-graph analysis and estimated coverage after applying the quality filter, providing acceptable sequence coverage for genome construction from short reads. Conclusions Quality filtering is a fundamental step in the process of constructing genomes, as it reduces the frequency of incorrect alignments that are caused by measuring errors, which can occur during the construction process due to the size of the reads, provoking misassemblies. Application of quality filters to sequence data, using the software Quality Assessment, along with graphing analyses, provided greater precision in the definition of cutoff parameters, which increased the accuracy of genome construction.

  10. Next-Generation Sequencing in Oncology: Genetic Diagnosis, Risk Prediction and Cancer Classification

    Directory of Open Access Journals (Sweden)

    Rick Kamps

    2017-01-01

    Full Text Available Next-generation sequencing (NGS technology has expanded in the last decades with significant improvements in the reliability, sequencing chemistry, pipeline analyses, data interpretation and costs. Such advances make the use of NGS feasible in clinical practice today. This review describes the recent technological developments in NGS applied to the field of oncology. A number of clinical applications are reviewed, i.e., mutation detection in inherited cancer syndromes based on DNA-sequencing, detection of spliceogenic variants based on RNA-sequencing, DNA-sequencing to identify risk modifiers and application for pre-implantation genetic diagnosis, cancer somatic mutation analysis, pharmacogenetics and liquid biopsy. Conclusive remarks, clinical limitations, implications and ethical considerations that relate to the different applications are provided.

  11. The complexity of sequences generated by the arc-fractal system.

    Science.gov (United States)

    Huynh, Hoai Nguyen; Pradana, Andri; Chew, Lock Yue

    2015-01-01

    We study properties of the symbolic sequences extracted from the fractals generated by the arc-fractal system introduced earlier by Huynh and Chew. The sequences consist of only a few symbols yet possess several nontrivial properties. First using an operator approach, we show that the sequences are not periodic, even though they are constructed from very simple rules. Second by employing the ϵ-machine approach developed by Crutchfield and Young, we measure the complexity and randomness of the sequences and show that they are indeed complex, i.e. neither periodic nor random, with the value of complexity measure being significant as compared to the known example of logistic map at the edge of chaos. The complexity and randomness of the sequences are then discussed in relation with the properties of associated fractal objects, such as their fractal dimension, symmetry and orientations of the arcs.

  12. The complexity of sequences generated by the arc-fractal system.

    Directory of Open Access Journals (Sweden)

    Hoai Nguyen Huynh

    Full Text Available We study properties of the symbolic sequences extracted from the fractals generated by the arc-fractal system introduced earlier by Huynh and Chew. The sequences consist of only a few symbols yet possess several nontrivial properties. First using an operator approach, we show that the sequences are not periodic, even though they are constructed from very simple rules. Second by employing the ϵ-machine approach developed by Crutchfield and Young, we measure the complexity and randomness of the sequences and show that they are indeed complex, i.e. neither periodic nor random, with the value of complexity measure being significant as compared to the known example of logistic map at the edge of chaos. The complexity and randomness of the sequences are then discussed in relation with the properties of associated fractal objects, such as their fractal dimension, symmetry and orientations of the arcs.

  13. Exploring the potential of second-generation sequencing in diverse biological contexts

    DEFF Research Database (Denmark)

    Fordyce, Sarah Louise

    Second generation sequencing (SGS) has revolutionized the study of DNA, allowing massive parallel sequencing of nucleic acids with unprecedented depths of coverage. The research undertaken in this thesis occurred in parallel with the increased accessibility of SGS platforms for routine genetic...... H1N1 influenza A virus genomes. The results of these studies demonstrate the power of SGS for gaining insight into the genetic variation of diverse biological samples and highlight the importance of using optimized protocols for sequencing non-conventional samples....

  14. Generation and Analysis of Full-length cDNA Sequences from Elephant Shark (Callorhinchus milii)

    KAUST Repository

    Kodzius, Rimantas

    2009-03-17

    Cartilaginous fishes are the oldest living group of jawed vertebrates and therefore is an important group for understanding the evolution of vertebrate genomes including the human genome. Our laboratory has proposed elephant shark (C. milii) as a model cartilaginous fish genome because of its relatively small genome size (910 Mb). The whole genome of C. milii is being sequenced (first cartilaginous fish genome to be sequenced completely). To characterize the transcriptome of C. milii and to assist in annotating exon-intron boundaries, transcriptional start sites and alternatively spliced transcripts, we are generating full-length cDNA sequences from C. milii.

  15. Genome wide SNP discovery in flax through next generation sequencing of reduced representation libraries

    Directory of Open Access Journals (Sweden)

    Kumar Santosh

    2012-12-01

    Full Text Available Abstract Background Flax (Linum usitatissimum L. is a significant fibre and oilseed crop. Current flax molecular markers, including isozymes, RAPDs, AFLPs and SSRs are of limited use in the construction of high density linkage maps and for association mapping applications due to factors such as low reproducibility, intense labour requirements and/or limited numbers. We report here on the use of a reduced representation library strategy combined with next generation Illumina sequencing for rapid and large scale discovery of SNPs in eight flax genotypes. SNP discovery was performed through in silico analysis of the sequencing data against the whole genome shotgun sequence assembly of flax genotype CDC Bethune. Genotyping-by-sequencing of an F6-derived recombinant inbred line population provided validation of the SNPs. Results Reduced representation libraries of eight flax genotypes were sequenced on the Illumina sequencing platform resulting in sequence coverage ranging from 4.33 to 15.64X (genome equivalents. Depending on the relatedness of the genotypes and the number and length of the reads, between 78% and 93% of the reads mapped onto the CDC Bethune whole genome shotgun sequence assembly. A total of 55,465 SNPs were discovered with the largest number of SNPs belonging to the genotypes with the highest mapping coverage percentage. Approximately 84% of the SNPs discovered were identified in a single genotype, 13% were shared between any two genotypes and the remaining 3% in three or more. Nearly a quarter of the SNPs were found in genic regions. A total of 4,706 out of 4,863 SNPs discovered in Macbeth were validated using genotyping-by-sequencing of 96 F6 individuals from a recombinant inbred line population derived from a cross between CDC Bethune and Macbeth, corresponding to a validation rate of 96.8%. Conclusions Next generation sequencing of reduced representation libraries was successfully implemented for genome-wide SNP discovery from

  16. Performance evaluation of the next-generation sequencing approach for molecular diagnosis of hereditary hearing loss.

    Science.gov (United States)

    Sivakumaran, Theru A; Husami, Ammar; Kissell, Diane; Zhang, Wenying; Keddache, Mehdi; Black, Angela P; Tinkle, Brad T; Greinwald, John H; Zhang, Kejian

    2013-06-01

    To evaluate the performance of a next-generation sequencing (NGS)-based targeted resequencing genetic test, OtoSeq, to identify the sequence variants in the genes causing sensorineural hearing loss (SNHL). Retrospective study. Tertiary children's hospital. A total of 8 individuals presenting with prelingual hearing loss were used in this study. The coding and flanking intronic regions of 24 well-studied SNHL genes were enriched using microdroplet polymerase chain reaction and sequenced on an Illumina HiSeq 2000 sequencer. The filtered high-quality sequence reads were mapped to reference sequence, and variants were detected using NextGENe software. A total of 1148 sequence variants were detected in 8 samples in 24 genes. Using in-house developed NGS data analysis criteria, we classified 810 (~71%) of these variants as potential true variants that include previously detected pathogenic mutations in 5 patients. To validate our strategy, we Sanger sequenced the target regions of 5 of the 24 genes, accounting for about 29.2% of all target sequence. Our results showed >99.99% concordance between NGS and Sanger sequencing in these 5 genes, resulting in an analytical sensitivity and specificity of 100% and 99.997%, respectively. We were able to successfully detect single base substitutions, small deletions, and insertions of up to 22 nucleotides. This study demonstrated that our NGS-based mutation screening strategy is highly sensitive and specific in detecting sequence variants in the SNHL genes. Therefore, we propose that this NGS-based targeted sequencing method would be an alternative to current technologies for identifying the multiple genetic causes of SNHL.

  17. Genome survey sequencing and genetic background characterization of Gracilariopsis lemaneiformis (Rhodophyta based on next-generation sequencing.

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    Wei Zhou

    Full Text Available Gracilariopsis lemaneiformis has a high economic value and is one of the most important aquaculture species in China. Despite it is economic importance, it has remained largely unstudied at the genomic level. In this study, we conducted a genome survey of Gp. lemaneiformis using next-generation sequencing (NGS technologies. In total, 18.70 Gb of high-quality sequence data with an estimated genome size of 97 Mb were obtained by HiSeq 2000 sequencing for Gp. lemaneiformis. These reads were assembled into 160,390 contigs with a N50 length of 3.64 kb, which were further assembled into 125,685 scaffolds with a total length of 81.17 Mb. Genome analysis predicted 3490 genes and a GC% content of 48%. The identified genes have an average transcript length of 1,429 bp, an average coding sequence size of 1,369 bp, 1.36 exons per gene, exon length of 1,008 bp, and intron length of 191 bp. From the initial assembled scaffold, transposable elements constituted 54.64% (44.35 Mb of the genome, and 7737 simple sequence repeats (SSRs were identified. Among these SSRs, the trinucleotide repeat type was the most abundant (up to 73.20% of total SSRs, followed by the di- (17.41%, tetra- (5.49%, hexa- (2.90%, and penta- (1.00% nucleotide repeat type. These characteristics suggest that Gp. lemaneiformis is a model organism for genetic study. This is the first report of genome-wide characterization within this taxon.

  18. A P-N Sequence Generator Using LFSR with Dual Edge Trigger Technique

    Directory of Open Access Journals (Sweden)

    Naghwal Nitin Kumar

    2016-01-01

    Full Text Available This paper represents the design and implementation of a low power 4-bit LFSR using Dual edge triggered flip flop. A linear feedback shift register (LFSR is assembled by N number of flip flops connected in series and a combinational logic generally xor gate. An LFSR can generate random number sequence which acts as cipher in cryptography. A known text encrypted over long PN sequence, in order to improve security sequence made longer ie 128 bit; require long chain of flip flop leads to more power consumption. In this paper a novel circuit of random sequence generator using dual edge triggered flip flop has been proposed. Data has been generated on every edge of flip flop instead of single edge. A DETFF-LFSR can generate random number require with less number of clock cycle, it minimizes the number of flip flop result in power saving. In this paper we concentrates on the designing of power competent Test Pattern Generator (TPG using four dual edge triggered flip-flops as the basic building block, overall there is reduction of power around 25% by using these techniques.

  19. Detection of Bacillus anthracis DNA in complex soil and air samples using next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Nicholas A Be

    Full Text Available Bacillus anthracis is the potentially lethal etiologic agent of anthrax disease, and is a significant concern in the realm of biodefense. One of the cornerstones of an effective biodefense strategy is the ability to detect infectious agents with a high degree of sensitivity and specificity in the context of a complex sample background. The nature of the B. anthracis genome, however, renders specific detection difficult, due to close homology with B. cereus and B. thuringiensis. We therefore elected to determine the efficacy of next-generation sequencing analysis and microarrays for detection of B. anthracis in an environmental background. We applied next-generation sequencing to titrated genome copy numbers of B. anthracis in the presence of background nucleic acid extracted from aerosol and soil samples. We found next-generation sequencing to be capable of detecting as few as 10 genomic equivalents of B. anthracis DNA per nanogram of background nucleic acid. Detection was accomplished by mapping reads to either a defined subset of reference genomes or to the full GenBank database. Moreover, sequence data obtained from B. anthracis could be reliably distinguished from sequence data mapping to either B. cereus or B. thuringiensis. We also demonstrated the efficacy of a microbial census microarray in detecting B. anthracis in the same samples, representing a cost-effective and high-throughput approach, complementary to next-generation sequencing. Our results, in combination with the capacity of sequencing for providing insights into the genomic characteristics of complex and novel organisms, suggest that these platforms should be considered important components of a biosurveillance strategy.

  20. Statistical inference of the generation probability of T-cell receptors from sequence repertoires.

    Science.gov (United States)

    Murugan, Anand; Mora, Thierry; Walczak, Aleksandra M; Callan, Curtis G

    2012-10-02

    Stochastic rearrangement of germline V-, D-, and J-genes to create variable coding sequence for certain cell surface receptors is at the origin of immune system diversity. This process, known as "VDJ recombination", is implemented via a series of stochastic molecular events involving gene choices and random nucleotide insertions between, and deletions from, genes. We use large sequence repertoires of the variable CDR3 region of human CD4+ T-cell receptor beta chains to infer the statistical properties of these basic biochemical events. Because any given CDR3 sequence can be produced in multiple ways, the probability distribution of hidden recombination events cannot be inferred directly from the observed sequences; we therefore develop a maximum likelihood inference method to achieve this end. To separate the properties of the molecular rearrangement mechanism from the effects of selection, we focus on nonproductive CDR3 sequences in T-cell DNA. We infer the joint distribution of the various generative events that occur when a new T-cell receptor gene is created. We find a rich picture of correlation (and absence thereof), providing insight into the molecular mechanisms involved. The generative event statistics are consistent between individuals, suggesting a universal biochemical process. Our probabilistic model predicts the generation probability of any specific CDR3 sequence by the primitive recombination process, allowing us to quantify the potential diversity of the T-cell repertoire and to understand why some sequences are shared between individuals. We argue that the use of formal statistical inference methods, of the kind presented in this paper, will be essential for quantitative understanding of the generation and evolution of diversity in the adaptive immune system.

  1. Recurrence Relations and Generating Functions of the Sequence of Sums of Corresponding Factorials and Triangular Numbers

    Directory of Open Access Journals (Sweden)

    Romer C. Castillo

    2015-11-01

    Full Text Available This study established some recurrence relations and exponential generating functions of the sequence of factoriangular numbers. A factoriangular number is defined as a sum of corresponding factorial and triangular number. The proofs utilize algebraic manipulations with some known results from calculus, particularly on power series and Maclaurin’s series. The recurrence relations were found by manipulating the formula defining a factoringular number while the ascertained exponential generating functions were in the closed form.

  2. Combined DECS Analysis and Next-Generation Sequencing Enable Efficient Detection of Novel Plant RNA Viruses.

    Science.gov (United States)

    Yanagisawa, Hironobu; Tomita, Reiko; Katsu, Koji; Uehara, Takuya; Atsumi, Go; Tateda, Chika; Kobayashi, Kappei; Sekine, Ken-Taro

    2016-03-07

    The presence of high molecular weight double-stranded RNA (dsRNA) within plant cells is an indicator of infection with RNA viruses as these possess genomic or replicative dsRNA. DECS (dsRNA isolation, exhaustive amplification, cloning, and sequencing) analysis has been shown to be capable of detecting unknown viruses. We postulated that a combination of DECS analysis and next-generation sequencing (NGS) would improve detection efficiency and usability of the technique. Here, we describe a model case in which we efficiently detected the presumed genome sequence of Blueberry shoestring virus (BSSV), a member of the genus Sobemovirus, which has not so far been reported. dsRNAs were isolated from BSSV-infected blueberry plants using the dsRNA-binding protein, reverse-transcribed, amplified, and sequenced using NGS. A contig of 4,020 nucleotides (nt) that shared similarities with sequences from other Sobemovirus species was obtained as a candidate of the BSSV genomic sequence. Reverse transcription (RT)-PCR primer sets based on sequences from this contig enabled the detection of BSSV in all BSSV-infected plants tested but not in healthy controls. A recombinant protein encoded by the putative coat protein gene was bound by the BSSV-antibody, indicating that the candidate sequence was that of BSSV itself. Our results suggest that a combination of DECS analysis and NGS, designated here as "DECS-C," is a powerful method for detecting novel plant viruses.

  3. Development of a low bias method for characterizing viral populations using next generation sequencing technology.

    Directory of Open Access Journals (Sweden)

    Stephanie M Willerth

    Full Text Available BACKGROUND: With an estimated 38 million people worldwide currently infected with human immunodeficiency virus (HIV, and an additional 4.1 million people becoming infected each year, it is important to understand how this virus mutates and develops resistance in order to design successful therapies. METHODOLOGY/PRINCIPAL FINDINGS: We report a novel experimental method for amplifying full-length HIV genomes without the use of sequence-specific primers for high throughput DNA sequencing, followed by assembly of full length viral genome sequences from the resulting large dataset. Illumina was chosen for sequencing due to its ability to provide greater coverage of the HIV genome compared to prior methods, allowing for more comprehensive characterization of the heterogeneity present in the HIV samples analyzed. Our novel amplification method in combination with Illumina sequencing was used to analyze two HIV populations: a homogenous HIV population based on the canonical NL4-3 strain and a heterogeneous viral population obtained from a HIV patient's infected T cells. In addition, the resulting sequence was analyzed using a new computational approach to obtain a consensus sequence and several metrics of diversity. SIGNIFICANCE: This study demonstrates how a lower bias amplification method in combination with next generation DNA sequencing provides in-depth, complete coverage of the HIV genome, enabling a stronger characterization of the quasispecies present in a clinically relevant HIV population as well as future study of how HIV mutates in response to a selective pressure.

  4. AMPLISAS: a web server for multilocus genotyping using next-generation amplicon sequencing data.

    Science.gov (United States)

    Sebastian, Alvaro; Herdegen, Magdalena; Migalska, Magdalena; Radwan, Jacek

    2016-03-01

    Next-generation sequencing (NGS) technologies are revolutionizing the fields of biology and medicine as powerful tools for amplicon sequencing (AS). Using combinations of primers and barcodes, it is possible to sequence targeted genomic regions with deep coverage for hundreds, even thousands, of individuals in a single experiment. This is extremely valuable for the genotyping of gene families in which locus-specific primers are often difficult to design, such as the major histocompatibility complex (MHC). The utility of AS is, however, limited by the high intrinsic sequencing error rates of NGS technologies and other sources of error such as polymerase amplification or chimera formation. Correcting these errors requires extensive bioinformatic post-processing of NGS data. Amplicon Sequence Assignment (AMPLISAS) is a tool that performs analysis of AS results in a simple and efficient way, while offering customization options for advanced users. AMPLISAS is designed as a three-step pipeline consisting of (i) read demultiplexing, (ii) unique sequence clustering and (iii) erroneous sequence filtering. Allele sequences and frequencies are retrieved in excel spreadsheet format, making them easy to interpret. AMPLISAS performance has been successfully benchmarked against previously published genotyped MHC data sets obtained with various NGS technologies.

  5. Fast and cost-effective genetic mapping in apple using next-generation sequencing.

    Science.gov (United States)

    Gardner, Kyle M; Brown, Patrick; Cooke, Thomas F; Cann, Scott; Costa, Fabrizio; Bustamante, Carlos; Velasco, Riccardo; Troggio, Michela; Myles, Sean

    2014-07-16

    Next-generation DNA sequencing (NGS) produces vast amounts of DNA sequence data, but it is not specifically designed to generate data suitable for genetic mapping. Recently developed DNA library preparation methods for NGS have helped solve this problem, however, by combining the use of reduced representation libraries with DNA sample barcoding to generate genome-wide genotype data from a common set of genetic markers across a large number of samples. Here we use such a method, called genotyping-by-sequencing (GBS), to produce a data set for genetic mapping in an F1 population of apples (Malus × domestica) segregating for skin color. We show that GBS produces a relatively large, but extremely sparse, genotype matrix: over 270,000 SNPs were discovered but most SNPs have too much missing data across samples to be useful for genetic mapping. After filtering for genotype quality and missing data, only 6% of the 85 million DNA sequence reads contributed to useful genotype calls. Despite this limitation, using existing software and a set of simple heuristics, we generated a final genotype matrix containing 3967 SNPs from 89 DNA samples from a single lane of Illumina HiSeq and used it to create a saturated genetic linkage map and to identify a known QTL underlying apple skin color. We therefore demonstrate that GBS is a cost-effective method for generating genome-wide SNP data suitable for genetic mapping in a highly diverse and heterozygous agricultural species. We anticipate future improvements to the GBS analysis pipeline presented here that will enhance the utility of next-generation DNA sequence data for the purposes of genetic mapping across diverse species.

  6. Generation of sequence-based data for pedigree-segregating Mendelian or Complex traits.

    Science.gov (United States)

    Li, Biao; Wang, Gao T; Leal, Suzanne M

    2015-11-15

    There is great interest in analyzing next generation sequence data that has been generated for pedigrees. However, unlike for population-based data there are only a limited number of rare variant methods to analyze pedigree data. One limitation is the ability to evaluate type I and II errors for family-based methods, due to lack of software that can simulate realistic sequence data for pedigrees. We developed RarePedSim (Rare-variant Pedigree-based Simulator), a program to simulate region/gene-level genotype and phenotype data for complex and Mendelian traits for any given pedigree structure. Using a genetic model, sequence variant data can be generated either conditionally or unconditionally on pedigree members' qualitative or quantitative phenotypes. Additionally, qualitative or quantitative traits can be generated conditional on variant data. Sequence data can either be simulated using realistic population demographic models or obtained from sequence-based studies. Variant sites can be annotated with positions, allele frequencies and functionality. For rare variants, RarePedSim is the only program that can efficiently generate both genotypes and phenotypes, regardless of pedigree structure. Data generated by RarePedSim are in standard Linkage file (.ped) and Variant Call (.vcf) formats, ready to be used for a variety of purposes, including evaluation of type I error and power, for association methods including mixed models and linkage analysis methods. bioinformatics.org/simped/rare sleal@bcm.edu. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  7. Generation and analysis of expressed sequence tags from the ciliate protozoan parasite Ichthyophthirius multifiliis

    Directory of Open Access Journals (Sweden)

    Arias Covadonga

    2007-06-01

    Full Text Available Abstract Background The ciliate protozoan Ichthyophthirius multifiliis (Ich is an important parasite of freshwater fish that causes 'white spot disease' leading to significant losses. A genomic resource for large-scale studies of this parasite has been lacking. To study gene expression involved in Ich pathogenesis and virulence, our goal was to generate expressed sequence tags (ESTs for the development of a powerful microarray platform for the analysis of global gene expression in this species. Here, we initiated a project to sequence and analyze over 10,000 ESTs. Results We sequenced 10,368 EST clones using a normalized cDNA library made from pooled samples of the trophont, tomont, and theront life-cycle stages, and generated 9,769 sequences (94.2% success rate. Post-sequencing processing led to 8,432 high quality sequences. Clustering analysis of these ESTs allowed identification of 4,706 unique sequences containing 976 contigs and 3,730 singletons. These unique sequences represent over two million base pairs (~10% of Plasmodium falciparum genome, a phylogenetically related protozoan. BLASTX searches produced 2,518 significant (E-value -5 hits and further Gene Ontology (GO analysis annotated 1,008 of these genes. The ESTs were analyzed comparatively against the genomes of the related protozoa Tetrahymena thermophila and P. falciparum, allowing putative identification of additional genes. All the EST sequences were deposited by dbEST in GenBank (GenBank: EG957858–EG966289. Gene discovery and annotations are presented and discussed. Conclusion This set of ESTs represents a significant proportion of the Ich transcriptome, and provides a material basis for the development of microarrays useful for gene expression studies concerning Ich development, pathogenesis, and virulence.

  8. Mitochondrial DNA variant discovery and evaluation in human Cardiomyopathies through next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Michael V Zaragoza

    Full Text Available Mutations in mitochondrial DNA (mtDNA may cause maternally-inherited cardiomyopathy and heart failure. In homoplasmy all mtDNA copies contain the mutation. In heteroplasmy there is a mixture of normal and mutant copies of mtDNA. The clinical phenotype of an affected individual depends on the type of genetic defect and the ratios of mutant and normal mtDNA in affected tissues. We aimed at determining the sensitivity of next-generation sequencing compared to Sanger sequencing for mutation detection in patients with mitochondrial cardiomyopathy. We studied 18 patients with mitochondrial cardiomyopathy and two with suspected mitochondrial disease. We "shotgun" sequenced PCR-amplified mtDNA and multiplexed using a single run on Roche's 454 Genome Sequencer. By mapping to the reference sequence, we obtained 1,300x average coverage per case and identified high-confidence variants. By comparing these to >400 mtDNA substitution variants detected by Sanger, we found 98% concordance in variant detection. Simulation studies showed that >95% of the homoplasmic variants were detected at a minimum sequence coverage of 20x while heteroplasmic variants required >200x coverage. Several Sanger "misses" were detected by 454 sequencing. These included the novel heteroplasmic 7501T>C in tRNA serine 1 in a patient with sudden cardiac death. These results support a potential role of next-generation sequencing in the discovery of novel mtDNA variants with heteroplasmy below the level reliably detected with Sanger sequencing. We hope that this will assist in the identification of mtDNA mutations and key genetic determinants for cardiomyopathy and mitochondrial disease.

  9. Characterization of mutations and sequence variants in the D21S11 locus by next generation sequencing

    DEFF Research Database (Denmark)

    Rockenbauer, Eszter; Hansen, Stine; Mikkelsen, Martin;

    2014-01-01

    We sequenced the D21S11 locus in 77 individuals from Danish paternity cases using 454 FLX next generation sequencing (NGS) technology. All samples were also typed with the AmpFlSTR(®) Profiler Plus(®) or the AmpFlSTR(®) Identifiler(®) PCR Amplification kits as part of paternity investigations....... In 18 of the confirmed trios, a genetic inconsistency was observed between one of the parents and the child at the D21S11 locus. NGS of the D21S11 locus revealed which allele had mutated from which parent to the child in 13 of these trios. All characterized mutations could be explained by single......-step mutations in the longest sub-repeat of D21S11. A total of 53 of the 77 sequenced samples originated from unrelated individuals. Twenty different D21S11 alleles were detected by NGS in these individuals whereas only 13 different alleles were observed with fragment analysis. Several alleles had the same...

  10. Multi-platform next-generation sequencing of the domestic turkey (Meleagris gallopavo) genome assembly and analysis

    Science.gov (United States)

    Next-generation sequencing technologies were used to rapidly and efficiently sequence the genome of the domestic turkey (Meleagris gallopavo). The current genome assembly (~1.1 Gb) includes 917 Mb of sequence assigned to chromosomes. Innate heterozygosity of the sequenced bird allowed discovery of...

  11. Challenges and opportunities in estimating viral genetic diversity from next-generation sequencing data

    Directory of Open Access Journals (Sweden)

    Niko eBeerenwinkel

    2012-09-01

    Full Text Available Many viruses, including the clinically relevant RNA viruses HIV and HCV, exist in large populations and display high genetic heterogeneity within and between infected hosts. Assessing intra-patient viral genetic diversity is essential for understanding the evolutionary dynamics of viruses, for designing effective vaccines, and for the success of antiviral therapy. Next-generation sequencing technologies allow the rapid and cost-effective acquisition of thousands to millions of short DNA sequences from a single sample. However, this approach entails several challenges in experimental design and computational data analysis. Here, we review the entire process of inferring viral diversity from sample collection to computing measures of genetic diversity. We discuss sample preparation, including reverse transcription and amplification, and the effect of experimental conditions on diversity estimates due to in vitro base substitutions, insertions, deletions, and recombination. The use of different next-generation sequencing platforms and their sequencing error profiles are compared in the context of various applications of diversity estimation, ranging from the detection of single nucleotide variants to the reconstruction of whole-genome haplotypes. We describe the statistical and computational challenges arising from these technical artifacts, and we review existing approaches, including available software, for their solution. Finally, we discuss open problems, and highlight successful biomedical applications and potential future clinical use of next-generation sequencing to estimate viral diversity.

  12. Generation and Analysis of Expressed Sequence Tags from Olea europaea L.

    Directory of Open Access Journals (Sweden)

    Nehir Ozdemir Ozgenturk

    2010-01-01

    Full Text Available Olive (Olea europaea L. is an important source of edible oil which was originated in Near-East region. In this study, two cDNA libraries were constructed from young olive leaves and immature olive fruits for generation of ESTs to discover the novel genes and search the function of unknown genes of olive. The randomly selected 3840 colonies were sequenced for EST collection from both libraries. Readable 2228 sequences for olive leaf and 1506 sequences for olive fruit were assembled into 205 and 69 contigs, respectively, whereas 2478 were singletons. Putative functions of all 2752 differentially expressed unique sequences were designated by gene homology based on BLAST and annotated using BLAST2GO. While 1339 ESTs show no homology to the database, 2024 ESTs have homology (under 80% with hypothetical proteins, putative proteins, expressed proteins, and unknown proteins in NCBI-GenBank. 635 EST's unique genes sequence have been identified by over 80% homology to known function in other species which were not previously described in Olea family. Only 3.1% of total EST's was shown similarity with olive database existing in NCBI. This generated EST's data and consensus sequences were submitted to NCBI as valuable source for functional genome studies of olive.

  13. Sarcomatoid adrenocortical carcinoma : a comprehensive pathological, immunohistochemical, and targeted next-generation sequencing analysis

    NARCIS (Netherlands)

    Papathomas, Thomas G.; Duregon, Eleonora; Korpershoek, Esther; Restuccia, David F.; van Marion, Ronald; Cappellesso, Rocco; Sturm, Nathalie; Rossi, Giulio; Coli, Antonella; Zucchini, Nicola; Stoop, Hans; Oosterhuis, Wolter; Ventura, Laura; Volante, Marco; Fassina, Ambrogio; Dinjens, Winand N M; Papotti, Mauro; de Krijger, Ronald R.

    2016-01-01

    Adrenocortical carcinomas (ACCs) with sarcomatous areas represent an extremely rare type of highly aggressive malignancy of unknown molecular pathogenesis. The current study was planned to gain insight into its molecular genetics using a targeted next-generation sequencing approach and to explore

  14. Body fluid identification of blood, saliva and semen using second generation sequencing of micro-RNA

    DEFF Research Database (Denmark)

    Petersen, Christel H.; Hjort, Benjamin Benn; Tvedebrink, Torben;

    2013-01-01

    We report a new second generation sequencing method for identification micro-RNA (miRNA) that can be used to identify body fluids and tissues. Principal component analysis of 10 miRNAs with high expression in 16 samples of blood, saliva and semen showed clear differences in the expression of mi...

  15. Body fluid identification of blood, saliva and semen using second generation sequencing of micro-RNA

    DEFF Research Database (Denmark)

    Petersen, Christel Hougård; Hjort, Benjamin Benn; Tvedebrink, Torben;

    2013-01-01

    We report a new second generation sequencing method for identification micro-RNA (miRNA) that can be used to identify body fluids and tissues. Principal component analysis of 10 miRNAs with high expression in 16 samples of blood, saliva and semen showed clear differences in the expression of miRN...

  16. Identification of Nucleotide Variation in Genomes Using Next-Generation Sequencing

    NARCIS (Netherlands)

    Megens, H.J.W.C.; Groenen, M.A.M.

    2012-01-01

    Discovery of genome-wide variation has taken a huge leap forward with the introduction of next-generation sequencing (NGS) technology. Variant discovery requires sampling of a number of haplotypes. This can be either the two haplotypes of a diploid organism or multiple haplotypes in a population. Va

  17. Scenario drafting for early technology assessment of next generation sequencing in clinical oncology

    NARCIS (Netherlands)

    Joosten, S.E.P.; Retel, V.P.; Coupé, V.M.H.; Heuvel, van den M.M.; Harten, van W.H.

    2016-01-01

    Background Next Generation Sequencing (NGS) is expected to lift molecular diagnostics in clinical oncology to the next level. It enables simultaneous identification of mutations in a patient tumor, after which targeted therapy may be assigned. This approach could improve patient survival and/or assi

  18. H-TArget model: Early technology assessment for ext generation sequencing in oncology

    NARCIS (Netherlands)

    Retel, Valesca P.; Joore, Manuela A.; Ramaekers, Bram; Heuvel, van den Michel M.; Heijden, van der Michiel Simon; Harten, van W.H.

    2015-01-01

    Background: Next Generation Sequencing (NGS) promises to find mutations (targets) in individual cancer patients, to subsequently prescribe targeted therapy. Currently, NGS is in development, the effects on choice of therapy and prognosis are still unclear, and the costs for targeted therapies are hi

  19. Unifying and generating of space vector modulation sequences for multilevel converter

    DEFF Research Database (Denmark)

    Ma, Ke; Blaabjerg, Frede

    2014-01-01

    Space Vector Modulation (SVM) is a powerful method which enables some freedom to generate the modulation sequences and modify the performances of converter. However, in the multi-level converter structures, the number of switching state redundancies significantly increases, and the determination...

  20. Efficient application of next-generation sequencing for the diagnosis of rare genetic syndromes

    NARCIS (Netherlands)

    Madrigal, I.; Alvarez-Mora, M.I.; Karlberg, O.; Rodriguez-Revenga, L.; Martinez Elurbe, D.; Rabionet, R.; Mur, A.; Pie, J.; Ballesta, F.; Sauer, S.; Syvanen, A.C.; Mila, M.

    2014-01-01

    AIMS: The causes of intellectual disability, which affects 1%-3% of the general population, are highly heterogeneous and the genetic defect remains unknown in around 40% of patients. The application of next-generation sequencing is changing the nature of biomedical diagnosis. This technology has qui

  1. Bringing Next-Generation Sequencing into the Classroom through a Comparison of Molecular Biology Techniques

    Science.gov (United States)

    Bowling, Bethany; Zimmer, Erin; Pyatt, Robert E.

    2014-01-01

    Although the development of next-generation (NextGen) sequencing technologies has revolutionized genomic research and medicine, the incorporation of these topics into the classroom is challenging, given an implied high degree of technical complexity. We developed an easy-to-implement, interactive classroom activity investigating the similarities…

  2. Sarcomatoid adrenocortical carcinoma : a comprehensive pathological, immunohistochemical, and targeted next-generation sequencing analysis

    NARCIS (Netherlands)

    Papathomas, Thomas G.; Duregon, Eleonora; Korpershoek, Esther; Restuccia, David F.; van Marion, Ronald; Cappellesso, Rocco; Sturm, Nathalie; Rossi, Giulio; Coli, Antonella; Zucchini, Nicola; Stoop, Hans; Oosterhuis, Wolter; Ventura, Laura; Volante, Marco; Fassina, Ambrogio; Dinjens, Winand N M; Papotti, Mauro; de Krijger, Ronald R.

    2016-01-01

    Adrenocortical carcinomas (ACCs) with sarcomatous areas represent an extremely rare type of highly aggressive malignancy of unknown molecular pathogenesis. The current study was planned to gain insight into its molecular genetics using a targeted next-generation sequencing approach and to explore th

  3. Internally generated sequences in learning and executing goal-directed behavior

    NARCIS (Netherlands)

    Pezzulo, G.; van der Meer, M.A.A.; Lansink, C.S.; Pennartz, C.M.A.

    2014-01-01

    A network of brain structures including hippocampus (HC), prefrontal cortex, and striatum controls goal-directed behavior and decision making. However, the neural mechanisms underlying these functions are unknown. Here, we review the role of 'internally generated sequences': structured, multi-neuron

  4. Next-generation sequencing for endocrine cancers: Recent advances and challenges.

    Science.gov (United States)

    Suresh, Padmanaban S; Venkatesh, Thejaswini; Tsutsumi, Rie; Shetty, Abhishek

    2017-05-01

    Contemporary molecular biology research tools have enriched numerous areas of biomedical research that address challenging diseases, including endocrine cancers (pituitary, thyroid, parathyroid, adrenal, testicular, ovarian, and neuroendocrine cancers). These tools have placed several intriguing clues before the scientific community. Endocrine cancers pose a major challenge in health care and research despite considerable attempts by researchers to understand their etiology. Microarray analyses have provided gene signatures from many cells, tissues, and organs that can differentiate healthy states from diseased ones, and even show patterns that correlate with stages of a disease. Microarray data can also elucidate the responses of endocrine tumors to therapeutic treatments. The rapid progress in next-generation sequencing methods has overcome many of the initial challenges of these technologies, and their advantages over microarray techniques have enabled them to emerge as valuable aids for clinical research applications (prognosis, identification of drug targets, etc.). A comprehensive review describing the recent advances in next-generation sequencing methods and their application in the evaluation of endocrine and endocrine-related cancers is lacking. The main purpose of this review is to illustrate the concepts that collectively constitute our current view of the possibilities offered by next-generation sequencing technological platforms, challenges to relevant applications, and perspectives on the future of clinical genetic testing of patients with endocrine tumors. We focus on recent discoveries in the use of next-generation sequencing methods for clinical diagnosis of endocrine tumors in patients and conclude with a discussion on persisting challenges and future objectives.

  5. Early detection of non-native fishes using next-generation DNA sequencing of fish larvae

    Science.gov (United States)

    Our objective was to evaluate the use of fish larvae for early detection of non-native fishes, comparing traditional and molecular taxonomy based on next-generation DNA sequencing to investigate potential efficiencies. Our approach was to intensively sample a Great Lakes non-nati...

  6. Efficient application of next-generation sequencing for the diagnosis of rare genetic syndromes

    NARCIS (Netherlands)

    Madrigal, I.; Alvarez-Mora, M.I.; Karlberg, O.; Rodriguez-Revenga, L.; Martinez Elurbe, D.; Rabionet, R.; Mur, A.; Pie, J.; Ballesta, F.; Sauer, S.; Syvanen, A.C.; Mila, M.

    2014-01-01

    AIMS: The causes of intellectual disability, which affects 1%-3% of the general population, are highly heterogeneous and the genetic defect remains unknown in around 40% of patients. The application of next-generation sequencing is changing the nature of biomedical diagnosis. This technology has qui

  7. Using next generation sequencing for multiplexed trait-linked markers in wheat

    Science.gov (United States)

    With the advent of next generation sequencing (NGS) technologies, single nucleotide polymorphisms (SNPs) have become the major type of marker for genotyping in many crops. However, the availability of SNP markers for important traits of bread wheat (Triticum aestivum L.) that can be effectively used...

  8. Karect: accurate correction of substitution, insertion and deletion errors for next-generation sequencing data

    KAUST Repository

    Allam, Amin

    2015-07-14

    Motivation: Next-generation sequencing generates large amounts of data affected by errors in the form of substitutions, insertions or deletions of bases. Error correction based on the high-coverage information, typically improves de novo assembly. Most existing tools can correct substitution errors only; some support insertions and deletions, but accuracy in many cases is low. Results: We present Karect, a novel error correction technique based on multiple alignment. Our approach supports substitution, insertion and deletion errors. It can handle non-uniform coverage as well as moderately covered areas of the sequenced genome. Experiments with data from Illumina, 454 FLX and Ion Torrent sequencing machines demonstrate that Karect is more accurate than previous methods, both in terms of correcting individual-bases errors (up to 10% increase in accuracy gain) and post de novo assembly quality (up to 10% increase in NGA50). We also introduce an improved framework for evaluating the quality of error correction.

  9. Quantitative miRNA expression analysis: comparing microarrays with next-generation sequencing

    DEFF Research Database (Denmark)

    Willenbrock, Hanni; Salomon, Jesper; Søkilde, Rolf

    2009-01-01

    Recently, next-generation sequencing has been introduced as a promising, new platform for assessing the copy number of transcripts, while the existing microarray technology is considered less reliable for absolute, quantitative expression measurements. Nonetheless, so far, results from the two...... technologies have only been compared based on biological data, leading to the conclusion that, although they are somewhat correlated, expression values differ significantly. Here, we use synthetic RNA samples, resembling human microRNA samples, to find that microarray expression measures actually correlate...... better with sample RNA content than expression measures obtained from sequencing data. In addition, microarrays appear highly sensitive and perform equivalently to next-generation sequencing in terms of reproducibility and relative ratio quantification....

  10. Next-generation sequencing reveals phylogeographic structure and a species tree for recent bird divergences

    DEFF Research Database (Denmark)

    McCormack, John E.; Maley, James M.; Hird, Sarah M.

    2012-01-01

    Next generation sequencing (NGS) technologies are revolutionizing many biological disciplines but have been slow to take root in phylogeography. This is partly due to the difficulty of using NGS to sequence orthologous DNA fragments for many individuals at low cost. We explore cases of recent...... divergence in four phylogenetically diverse avian systems using a method for quick and cost-effective generation of primary DNA sequence data using pyrosequencing. NGS data were processed using an analytical pipeline that reduces many reads into two called alleles per locus per individual. Using single...... nucleotide polymorphisms (SNPs) mined from the loci, we detected population differentiation in each of the four bird systems, including: a case of ecological speciation in rails (Rallus); a rapid postglacial radiation in the genus Junco; recent in situ speciation among hummingbirds (Trochilus) in Jamaica...

  11. Perspectives of DNA microarray and next-generation DNA sequencing technologies

    Institute of Scientific and Technical Information of China (English)

    TENG XiaoKun; XIAO HuaSheng

    2009-01-01

    DNA microarray and next-generation DNA sequencing technologies are important tools for high-throughput genome research, in revealing both the structural and functional characteristics of genomes. In the past decade the DNA microarray technologies have been widely applied in the studies of functional genomics, systems biology and pharmacogenomics. The next-generation DNA sequenc-ing method was first introduced by the 454 Company in 2003, immediately followed by the establish-ment of the Solexa and Solid techniques by other biotech companies. Though it has not been long since the first emergence of this technology, with the fast and impressive improvement, the application of this technology has extended to almost all fields of genomics research, as a rival challenging the existing DNA microarray technology. This paper briefly reviews the working principles of these two technologies as well as their application and perspectives in genome research.

  12. Counting of oligomers in sequences generated by markov chains for DNA motif discovery.

    Science.gov (United States)

    Shan, Gao; Zheng, Wei-Mou

    2009-02-01

    By means of the technique of the imbedded Markov chain, an efficient algorithm is proposed to exactly calculate first, second moments of word counts and the probability for a word to occur at least once in random texts generated by a Markov chain. A generating function is introduced directly from the imbedded Markov chain to derive asymptotic approximations for the problem. Two Z-scores, one based on the number of sequences with hits and the other on the total number of word hits in a set of sequences, are examined for discovery of motifs on a set of promoter sequences extracted from A. thaliana genome. Source code is available at http://www.itp.ac.cn/zheng/oligo.c.

  13. Fiscal 1995 survey of promotion of the geothermal development. Report on a usage feasibility test of a small scale geothermal binary cycle power generation system; 1995 nendo chinetsu kaihatsu sokushin chosa. Chusho chinetsu binary hatsuden system jissho shiken hokokusho

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-03-01

    In this survey, studies for popularization and practical utilization of small and medium size geothermal binary cycle power systems which assesses low and medium temperature geothermal resources were conducted, and studies for development of the system to be introduced for practical use and for promotion of the popularization were made. A study was carried out of preconditions and various conditions of a demonstrative test plant (100kW class, 500kW class) in view of the initial cost of the actual plant, and an analysis was made of the power generation cost. Acceptability of the demonstrative test plant (100kW class) was examined to analyze problems on the introduction. A thermodynamic analysis was made of the output of geothermal binary cycle power generation. Analysis/evaluation of the results of the 100kW demonstrative test plant were carried out in view of the operation results of the plant of the same kind, and checks/reviews were conducted of performance and reliability of the system, equipment simplification, etc. Inspection of the system was made in the stage of design/manufacture of the 500kW demonstrative test plant. Concerning the spread/expansion of the system, studied were multiple stage geothermal utilization and PR promotion method. 14 refs., 62 figs., 55 tabs.

  14. Partial sequence homogenization in the 5S multigene families may generate sequence chimeras and spurious results in phylogenetic reconstructions.

    Science.gov (United States)

    Galián, José A; Rosato, Marcela; Rosselló, Josep A

    2014-03-01

    Multigene families have provided opportunities for evolutionary biologists to assess molecular evolution processes and phylogenetic reconstructions at deep and shallow systematic levels. However, the use of these markers is not free of technical and analytical challenges. Many evolutionary studies that used the nuclear 5S rDNA gene family rarely used contiguous 5S coding sequences due to the routine use of head-to-tail polymerase chain reaction primers that are anchored to the coding region. Moreover, the 5S coding sequences have been concatenated with independent, adjacent gene units in many studies, creating simulated chimeric genes as the raw data for evolutionary analysis. This practice is based on the tacitly assumed, but rarely tested, hypothesis that strict intra-locus concerted evolution processes are operating in 5S rDNA genes, without any empirical evidence as to whether it holds for the recovered data. The potential pitfalls of analysing the patterns of molecular evolution and reconstructing phylogenies based on these chimeric genes have not been assessed to date. Here, we compared the sequence integrity and phylogenetic behavior of entire versus concatenated 5S coding regions from a real data set obtained from closely related plant species (Medicago, Fabaceae). Our results suggest that within arrays sequence homogenization is partially operating in the 5S coding region, which is traditionally assumed to be highly conserved. Consequently, concatenating 5S genes increases haplotype diversity, generating novel chimeric genotypes that most likely do not exist within the genome. In addition, the patterns of gene evolution are distorted, leading to incorrect haplotype relationships in some evolutionary reconstructions.

  15. Development and evaluation of a panel of filovirus sequence capture probes for pathogen detection by next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Jeffrey W Koehler

    Full Text Available A detailed understanding of the circulating pathogens in a particular geographic location aids in effectively utilizing targeted, rapid diagnostic assays, thus allowing for appropriate therapeutic and containment procedures. This is especially important in regions prevalent for highly pathogenic viruses co-circulating with other endemic pathogens such as the malaria parasite. The importance of biosurveillance is highlighted by the ongoing Ebola virus disease outbreak in West Africa. For example, a more comprehensive assessment of the regional pathogens could have identified the risk of a filovirus disease outbreak earlier and led to an improved diagnostic and response capacity in the region. In this context, being able to rapidly screen a single sample for multiple pathogens in a single tube reaction could improve both diagnostics as well as pathogen surveillance. Here, probes were designed to capture identifying filovirus sequence for the ebolaviruses Sudan, Ebola, Reston, Taï Forest, and Bundibugyo and the Marburg virus variants Musoke, Ci67, and Angola. These probes were combined into a single probe panel, and the captured filovirus sequence was successfully identified using the MiSeq next-generation sequencing platform. This panel was then used to identify the specific filovirus from nonhuman primates experimentally infected with Ebola virus as well as Bundibugyo virus in human sera samples from the Democratic Republic of the Congo, thus demonstrating the utility for pathogen detection using clinical samples. While not as sensitive and rapid as real-time PCR, this panel, along with incorporating additional sequence capture probe panels, could be used for broad pathogen screening and biosurveillance.

  16. Z-DNA-forming sequences generate large-scale deletions in mammalian cells.

    Science.gov (United States)

    Wang, Guliang; Christensen, Laura A; Vasquez, Karen M

    2006-02-21

    Spontaneous chromosomal breakages frequently occur at genomic hot spots in the absence of DNA damage and can result in translocation-related human disease. Chromosomal breakpoints are often mapped near purine-pyrimidine Z-DNA-forming sequences in human tumors. However, it is not known whether Z-DNA plays a role in the generation of these chromosomal breakages. Here, we show that Z-DNA-forming sequences induce high levels of genetic instability in both bacterial and mammalian cells. In mammalian cells, the Z-DNA-forming sequences induce double-strand breaks nearby, resulting in large-scale deletions in 95% of the mutants. These Z-DNA-induced double-strand breaks in mammalian cells are not confined to a specific sequence but rather are dispersed over a 400-bp region, consistent with chromosomal breakpoints in human diseases. This observation is in contrast to the mutations generated in Escherichia coli that are predominantly small deletions within the repeats. We found that the frequency of small deletions is increased by replication in mammalian cell extracts. Surprisingly, the large-scale deletions generated in mammalian cells are, at least in part, replication-independent and are likely initiated by repair processing cleavages surrounding the Z-DNA-forming sequence. These results reveal that mammalian cells process Z-DNA-forming sequences in a strikingly different fashion from that used by bacteria. Our data suggest that Z-DNA-forming sequences may be causative factors for gene translocations found in leukemias and lymphomas and that certain cellular conditions such as active transcription may increase the risk of Z-DNA-related genetic instability.

  17. Next-generation sequencing technologies: breaking the sound barrier of human genetics.

    Science.gov (United States)

    Bahassi, El Mustapha; Stambrook, Peter J

    2014-09-01

    Demand for new technologies that deliver fast, inexpensive and accurate genome information has never been greater. This challenge has catalysed the rapid development of advances in next-generation sequencing (NGS). The generation of large volumes of sequence data and the speed of data acquisition are the primary advantages over previous, more standard methods. In 2013, the Food and Drug Administration granted marketing authorisation for the first high-throughput NG sequencer, Illumina's MiSeqDx, which allowed the development and use of a large number of new genome-based tests. Here, we present a review of template preparation, nucleic acid sequencing and imaging, genome assembly and alignment approaches as well as recent advances in current and near-term commercially available NGS instruments. We also outline the broad range of applications for NGS technologies and provide guidelines for platform selection to best address biological questions of interest. DNA sequencing has revolutionised biological and medical research, and is poised to have a similar impact on the practice of medicine. This tool is but one of an increasing arsenal of developing tools that enhance our capabilities to identify, quantify and functionally characterise the components of biological networks that keep us healthy or make us sick. Despite advances in other 'omic' technologies, DNA sequencing and analysis, in many respects, have played the leading role to date. The new technologies provide a bridge between genotype and phenotype, both in man and model organisms, and have revolutionised how risk of developing a complex human disease may be assessed. The generation of large DNA sequence data sets is producing a wealth of medically relevant information on a large number of individuals and populations that will potentially form the basis of truly individualised medical care in the future.

  18. Chaotic-Laser-Based True Random Sequence Generation for Spread-Spectrum Communications

    Institute of Scientific and Technical Information of China (English)

    张朝霞; 周俊杰; 张东泽; 傅正; 张建忠

    2012-01-01

    We propose a scheme for spread-spectrum communications using true random sequences generated by chaotic semiconductor lasers as spreading codes. These sequences can eliminate the inherent periodicity of pseudorandom sequences,enlarge the capacity of spread-spectrum codes,improve communication security,and increase the number ot users of the system.When a true random sequence with an appropriate length is used as thespread-spectrum code and the information speed is maintained constant,the system acquires a greater spreadspectrum gain and a lower bit-error ratio (BER) than the traditional spread-spectrum system.The communication security is also enhanced.The BER smoothly increases with the number of users,which indicates the good multipleaccess capability of the system.

  19. Considerations for clinical read alignment and mutational profiling using next-generation sequencing

    Directory of Open Access Journals (Sweden)

    Gavin R Oliver

    2012-07-01

    Full Text Available Next-generation sequencing technologies are increasingly being applied in clinical settings, however the data are characterized by a range of platform-specific artifacts making downstream analysis problematic and error prone. One major application of NGS is in the profiling of clinically relevant mutations whereby sequences are aligned to a reference genome and potential mutations assessed and scored. Accurate sequence alignment is pivotal in reliable assessment of potential mutations however selection of appropriate alignment tools is a non-trivial task complicated by the availability of multiple solutions each with its own performance characteristics. Using BRCA1 as an example, we have simulated and mutated a test dataset based on Illumina sequencing technology. Our findings reveal key differences in the performances of a range of common commercial and open source tools and will be of importance to anyone using NGS to profile mutations in clinical or basic research.

  20. Next-generation sequencing and metagenomic analysis: a universal diagnostic tool in plant virology.

    Science.gov (United States)

    Adams, Ian P; Glover, Rachel H; Monger, Wendy A; Mumford, Rick; Jackeviciene, Elena; Navalinskiene, Meletele; Samuitiene, Marija; Boonham, Neil

    2009-07-01

    A novel, unbiased approach to plant viral disease diagnosis has been developed which requires no a priori knowledge of the host or pathogen. Next-generation sequencing coupled with metagenomic analysis was used to produce large quantities of cDNA sequence in a model system of tomato infected with Pepino mosaic virus. The method was then applied to a sample of Gomphrena globosa infected with an unknown pathogen originally isolated from the flowering plant Liatris spicata. This plant was found to contain a new cucumovirus, for which we suggest the name 'Gayfeather mild mottle virus'. In both cases, the full viral genome was sequenced. This method expedites the entire process of novel virus discovery, identification, viral genome sequencing and, subsequently, the development of more routine assays for new viral pathogens.

  1. DNA fingerprinting, DNA barcoding, and next generation sequencing technology in plants.

    Science.gov (United States)

    Sucher, Nikolaus J; Hennell, James R; Carles, Maria C

    2012-01-01

    DNA fingerprinting of plants has become an invaluable tool in forensic, scientific, and industrial laboratories all over the world. PCR has become part of virtually every variation of the plethora of approaches used for DNA fingerprinting today. DNA sequencing is increasingly used either in combination with or as a replacement for traditional DNA fingerprinting techniques. A prime example is the use of short, standardized regions of the genome as taxon barcodes for biological identification of plants. Rapid advances in "next generation sequencing" (NGS) technology are driving down the cost of sequencing and bringing large-scale sequencing projects into the reach of individual investigators. We present an overview of recent publications that demonstrate the use of "NGS" technology for DNA fingerprinting and DNA barcoding applications.

  2. Next-generation sequencing technology for genetics and genomics of sorghum

    DEFF Research Database (Denmark)

    Luo, Hong; Mocoeur, Anne Raymonde Joelle; Jing, Hai-Chun

    2014-01-01

    NGS platforms, comparing their working theories and reveiwing their advantages and disavantages. We also discuss the future of NGS development and point out that single molecular sequencing would push the technology to the next level for biological sciences. Much of the chapter focuses on the use......The invention and application of Next-Generation Sequencing (NGS) technologies have revolutionized the study of genetics and genomics. Much research which would not even be considered are nowdays being excuted in many laboratories as routine. In this chapter, we introduce the currently available...... of NGS technologies in sorghum. Although the acquisition of the first whole-genome sequence in sorghum was carried out primarily using Sanger sequencing, the use of NGS for examining the genome-wide variation was almost synchronized with other work. Interesting genomic variation was found between sweet...

  3. A Primer for Disease Gene Prioritization Using Next-Generation Sequencing Data

    Directory of Open Access Journals (Sweden)

    Shuoguo Wang

    2013-12-01

    Full Text Available High-throughput next-generation sequencing (NGS technology produces a tremendous amount of raw sequence data. The challenges for researchers are to process the raw data, to map the sequences to genome, to discover variants that are different from the reference genome, and to prioritize/rank the variants for the question of interest. The recent development of many computational algorithms and programs has vastly improved the ability to translate sequence data into valuable information for disease gene identification. However, the NGS data analysis is complex and could be overwhelming for researchers who are not familiar with the process. Here, we outline the analysis pipeline and describe some of the most commonly used principles and tools for analyzing NGS data for disease gene identification.

  4. SIS: a program to generate draft genome sequence scaffolds for prokaryotes

    Directory of Open Access Journals (Sweden)

    Dias Zanoni

    2012-05-01

    Full Text Available Abstract Background Decreasing costs of DNA sequencing have made prokaryotic draft genome sequences increasingly common. A contig scaffold is an ordering of contigs in the correct orientation. A scaffold can help genome comparisons and guide gap closure efforts. One popular technique for obtaining contig scaffolds is to map contigs onto a reference genome. However, rearrangements that may exist between the query and reference genomes may result in incorrect scaffolds, if these rearrangements are not taken into account. Large-scale inversions are common rearrangement events in prokaryotic genomes. Even in draft genomes it is possible to detect the presence of inversions given sufficient sequencing coverage and a sufficiently close reference genome. Results We present a linear-time algorithm that can generate a set of contig scaffolds for a draft genome sequence represented in contigs given a reference genome. The algorithm is aimed at prokaryotic genomes and relies on the presence of matching sequence patterns between the query and reference genomes that can be interpreted as the result of large-scale inversions; we call these patterns inversion signatures. Our algorithm is capable of correctly generating a scaffold if at least one member of every inversion signature pair is present in contigs and no inversion signatures have been overwritten in evolution. The algorithm is also capable of generating scaffolds in the presence of any kind of inversion, even though in this general case there is no guarantee that all scaffolds in the scaffold set will be correct. We compare the performance of sis, the program that implements the algorithm, to seven other scaffold-generating programs. The results of our tests show that sis has overall better performance. Conclusions sis is a new easy-to-use tool to generate contig scaffolds, available both as stand-alone and as a web server. The good performance of sis in our tests adds evidence that large

  5. Very high resolution single pass HLA genotyping using amplicon sequencing on the 454 next generation DNA sequencers: Comparison with Sanger sequencing.

    Science.gov (United States)

    Yamamoto, F; Höglund, B; Fernandez-Vina, M; Tyan, D; Rastrou, M; Williams, T; Moonsamy, P; Goodridge, D; Anderson, M; Erlich, H A; Holcomb, C L

    2015-12-01

    Compared to Sanger sequencing, next-generation sequencing offers advantages for high resolution HLA genotyping including increased throughput, lower cost, and reduced genotype ambiguity. Here we describe an enhancement of the Roche 454 GS GType HLA genotyping assay to provide very high resolution (VHR) typing, by the addition of 8 primer pairs to the original 14, to genotype 11 HLA loci. These additional amplicons help resolve common and well-documented alleles and exclude commonly found null alleles in genotype ambiguity strings. Simplification of workflow to reduce the initial preparation effort using early pooling of amplicons or the Fluidigm Access Array™ is also described. Performance of the VHR assay was evaluated on 28 well characterized cell lines using Conexio Assign MPS software which uses genomic, rather than cDNA, reference sequence. Concordance was 98.4%; 1.6% had no genotype assignment. Of concordant calls, 53% were unambiguous. To further assess the assay, 59 clinical samples were genotyped and results compared to unambiguous allele assignments obtained by prior sequence-based typing supplemented with SSO and/or SSP. Concordance was 98.7% with 58.2% as unambiguous calls; 1.3% could not be assigned. Our results show that the amplicon-based VHR assay is robust and can replace current Sanger methodology. Together with software enhancements, it has the potential to provide even higher resolution HLA typing. Copyright © 2015. Published by Elsevier Inc.

  6. NGS-QC Generator: A Quality Control System for ChIP-Seq and Related Deep Sequencing-Generated Datasets.

    Science.gov (United States)

    Mendoza-Parra, Marco Antonio; Saleem, Mohamed-Ashick M; Blum, Matthias; Cholley, Pierre-Etienne; Gronemeyer, Hinrich

    2016-01-01

    The combination of massive parallel sequencing with a variety of modern DNA/RNA enrichment technologies provides means for interrogating functional protein-genome interactions (ChIP-seq), genome-wide transcriptional activity (RNA-seq; GRO-seq), chromatin accessibility (DNase-seq, FAIRE-seq, MNase-seq), and more recently the three-dimensional organization of chromatin (Hi-C, ChIA-PET). In systems biology-based approaches several of these readouts are generally cumulated with the aim of describing living systems through a reconstitution of the genome-regulatory functions. However, an issue that is often underestimated is that conclusions drawn from such multidimensional analyses of NGS-derived datasets critically depend on the quality of the compared datasets. To address this problem, we have developed the NGS-QC Generator, a quality control system that infers quality descriptors for any kind of ChIP-sequencing and related datasets. In this chapter we provide a detailed protocol for (1) assessing quality descriptors with the NGS-QC Generator; (2) to interpret the generated reports; and (3) to explore the database of QC indicators (www.ngs-qc.org) for >21,000 publicly available datasets.

  7. Bioinformatic Challenges in Clinical Diagnostic Application of Targeted Next Generation Sequencing: Experience from Pheochromocytoma.

    Directory of Open Access Journals (Sweden)

    Joakim Crona

    Full Text Available Recent studies have demonstrated equal quality of targeted next generation sequencing (NGS compared to Sanger Sequencing. Whereas these novel sequencing processes have a validated robust performance, choice of enrichment method and different available bioinformatic software as reliable analysis tool needs to be further investigated in a diagnostic setting.DNA from 21 patients with genetic variants in SDHB, VHL, EPAS1, RET, (n=17 or clinical criteria of NF1 syndrome (n=4 were included. Targeted NGS was performed using Truseq custom amplicon enrichment sequenced on an Illumina MiSEQ instrument. Results were analysed in parallel using three different bioinformatics pipelines; (1 Commercially available MiSEQ Reporter, fully automatized and integrated software, (2 CLC Genomics Workbench, graphical interface based software, also commercially available, and ICP (3 an in-house scripted custom bioinformatic tool.A tenfold read coverage was achieved in between 95-98% of targeted bases. All workflows had alignment of reads to SDHA and NF1 pseudogenes. Compared to Sanger sequencing, variant calling revealed a sensitivity ranging from 83 to 100% and a specificity of 99.9-100%. Only MiSEQ reporter identified all pathogenic variants in both sequencing runs.We conclude that targeted next generation sequencing have equal quality compared to Sanger sequencing. Enrichment specificity and the bioinformatic performance need to be carefully assessed in a diagnostic setting. As acceptable accuracy was noted for a fully automated bioinformatic workflow, we suggest that processing of NGS data could be performed without expert bioinformatics skills utilizing already existing commercially available bioinformatics tools.

  8. Computel: computation of mean telomere length from whole-genome next-generation sequencing data.

    Science.gov (United States)

    Nersisyan, Lilit; Arakelyan, Arsen

    2015-01-01

    Telomeres are the ends of eukaryotic chromosomes, consisting of consecutive short repeats that protect chromosome ends from degradation. Telomeres shorten with each cell division, leading to replicative cell senescence. Deregulation of telomere length homeostasis is associated with the development of various age-related diseases and cancers. A number of experimental techniques exist for telomere length measurement; however, until recently, the absence of tools for extracting telomere lengths from high-throughput sequencing data has significantly obscured the association of telomere length with molecular processes in normal and diseased conditions. We have developed Computel, a program in R for computing mean telomere length from whole-genome next-generation sequencing data. Computel is open source, and is freely available at https://github.com/lilit-nersisyan/computel. It utilizes a short-read alignment-based approach and integrates various popular tools for sequencing data analysis. We validated it with synthetic and experimental data, and compared its performance with the previously available software. The results have shown that Computel outperforms existing software in accuracy, independence of results from sequencing conditions, stability against inherent sequencing errors, and better ability to distinguish pure telomeric sequences from interstitial telomeric repeats. By providing a highly reliable methodology for determining telomere lengths from whole-genome sequencing data, Computel should help to elucidate the role of telomeres in cellular health and disease.

  9. Computel: computation of mean telomere length from whole-genome next-generation sequencing data.

    Directory of Open Access Journals (Sweden)

    Lilit Nersisyan

    Full Text Available Telomeres are the ends of eukaryotic chromosomes, consisting of consecutive short repeats that protect chromosome ends from degradation. Telomeres shorten with each cell division, leading to replicative cell senescence. Deregulation of telomere length homeostasis is associated with the development of various age-related diseases and cancers. A number of experimental techniques exist for telomere length measurement; however, until recently, the absence of tools for extracting telomere lengths from high-throughput sequencing data has significantly obscured the association of telomere length with molecular processes in normal and diseased conditions. We have developed Computel, a program in R for computing mean telomere length from whole-genome next-generation sequencing data. Computel is open source, and is freely available at https://github.com/lilit-nersisyan/computel. It utilizes a short-read alignment-based approach and integrates various popular tools for sequencing data analysis. We validated it with synthetic and experimental data, and compared its performance with the previously available software. The results have shown that Computel outperforms existing software in accuracy, independence of results from sequencing conditions, stability against inherent sequencing errors, and better ability to distinguish pure telomeric sequences from interstitial telomeric repeats. By providing a highly reliable methodology for determining telomere lengths from whole-genome sequencing data, Computel should help to elucidate the role of telomeres in cellular health and disease.

  10. Development of multidimensional sequence operation theory with applications to risk evaluation in power system generation scheduling

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Sequence operation theory (SOT) is a powerful tool for solving complex probabil-istic problems in power system. However, the basic single dimension SOT cannot satisfy the requirement of multi-state and multi-attribute analysis, which is often the case in actual power system practice. To address this problem, multidimensional sequence operation theory (MSOT) is developed. On the basis of previous research, this paper first categorizes the situations by the number of state variables and the number of attribute values, and defines the multidimensional sequence for single state variable and multiple attribute values, as well as the multidimensional se-quence for multiple state variables and multiple attribute values. Corresponding to those definitions, four types of operations between two discrete multidimensional sequences are derived respectively. Therefore, the sequence is extended from sin-gle dimensional to multidimensional, establishing an integrated theory of multidi-mensional sequence operation. In particular, the basic single dimension SOT can be viewed as a special case of MSOT with only one state variable and one attribute value. Finally, the paper demonstrates the effectiveness of MSOT through an ex-ample of risk evaluation in power system generation scheduling.

  11. Comparing Apples and Oranges?: Next Generation Sequencing and Its Impact on Microbiome Analysis.

    Directory of Open Access Journals (Sweden)

    Adam G Clooney

    Full Text Available Rapid advancements in sequencing technologies along with falling costs present widespread opportunities for microbiome studies across a vast and diverse array of environments. These impressive technological developments have been accompanied by a considerable growth in the number of methodological variables, including sampling, storage, DNA extraction, primer pairs, sequencing technology, chemistry version, read length, insert size, and analysis pipelines, amongst others. This increase in variability threatens to compromise both the reproducibility and the comparability of studies conducted. Here we perform the first reported study comparing both amplicon and shotgun sequencing for the three leading next-generation sequencing technologies. These were applied to six human stool samples using Illumina HiSeq, MiSeq and Ion PGM shotgun sequencing, as well as amplicon sequencing across two variable 16S rRNA gene regions. Notably, we found that the factor responsible for the greatest variance in microbiota composition was the chosen methodology rather than the natural inter-individual variance, which is commonly one of the most significant drivers in microbiome studies. Amplicon sequencing suffered from this to a large extent, and this issue was particularly apparent when the 16S rRNA V1-V2 region amplicons were sequenced with MiSeq. Somewhat surprisingly, the choice of taxonomic binning software for shotgun sequences proved to be of crucial importance with even greater discriminatory power than sequencing technology and choice of amplicon. Optimal N50 assembly values for the HiSeq was obtained for 10 million reads per sample, whereas the applied MiSeq and PGM sequencing depths proved less sufficient for shotgun sequencing of stool samples. The latter technologies, on the other hand, provide a better basis for functional gene categorisation, possibly due to their longer read lengths. Hence, in addition to highlighting methodological biases, this study

  12. Comparing Apples and Oranges?: Next Generation Sequencing and Its Impact on Microbiome Analysis.

    Science.gov (United States)

    Clooney, Adam G; Fouhy, Fiona; Sleator, Roy D; O' Driscoll, Aisling; Stanton, Catherine; Cotter, Paul D; Claesson, Marcus J

    2016-01-01

    Rapid advancements in sequencing technologies along with falling costs present widespread opportunities for microbiome studies across a vast and diverse array of environments. These impressive technological developments have been accompanied by a considerable growth in the number of methodological variables, including sampling, storage, DNA extraction, primer pairs, sequencing technology, chemistry version, read length, insert size, and analysis pipelines, amongst others. This increase in variability threatens to compromise both the reproducibility and the comparability of studies conducted. Here we perform the first reported study comparing both amplicon and shotgun sequencing for the three leading next-generation sequencing technologies. These were applied to six human stool samples using Illumina HiSeq, MiSeq and Ion PGM shotgun sequencing, as well as amplicon sequencing across two variable 16S rRNA gene regions. Notably, we found that the factor responsible for the greatest variance in microbiota composition was the chosen methodology rather than the natural inter-individual variance, which is commonly one of the most significant drivers in microbiome studies. Amplicon sequencing suffered from this to a large extent, and this issue was particularly apparent when the 16S rRNA V1-V2 region amplicons were sequenced with MiSeq. Somewhat surprisingly, the choice of taxonomic binning software for shotgun sequences proved to be of crucial importance with even greater discriminatory power than sequencing technology and choice of amplicon. Optimal N50 assembly values for the HiSeq was obtained for 10 million reads per sample, whereas the applied MiSeq and PGM sequencing depths proved less sufficient for shotgun sequencing of stool samples. The latter technologies, on the other hand, provide a better basis for functional gene categorisation, possibly due to their longer read lengths. Hence, in addition to highlighting methodological biases, this study demonstrates the

  13. Pulse Sequences for Efficient Multi-Cycle Terahertz Generation in Periodically Poled Lithium Niobate

    CERN Document Server

    Ravi, Koustuban; Kärtner, Franz X

    2016-01-01

    The use of laser pulse sequences to drive the cascaded difference frequency generation of high energy, high peak-power and multi-cycle terahertz pulses in cryogenically cooled periodically poled lithium niobate is proposed. Detailed simulations considering the coupled nonlinear interaction of terahertz and optical waves show that unprecedented optical-to-terahertz energy conversion efficiencies > 5%, peak electric fields of hundred(s) of Mega volts/meter at terahertz pulse durations of hundred(s) of picoseconds can be achieved. The proposed methods are shown to circumvent laser-induced damage at Joule-level pumping by 1$\\mu$m lasers to enable multi-cycle terahertz sources with pulse energies >> 10 milli-joules. Various pulse sequence formats are proposed and analyzed. Numerical calculations for periodically poled structures accounting for cascaded difference frequency generation, self-phase-modulation, cascaded second harmonic generation and laser induced damage are introduced. Unprecedented studies of the ph...

  14. Rapid Detection of Rare Deleterious Variants by Next Generation Sequencing with Optional Microarray SNP Genotype Data.

    Science.gov (United States)

    Watson, Christopher M; Crinnion, Laura A; Gurgel-Gianetti, Juliana; Harrison, Sally M; Daly, Catherine; Antanavicuite, Agne; Lascelles, Carolina; Markham, Alexander F; Pena, Sergio D J; Bonthron, David T; Carr, Ian M

    2015-09-01

    Autozygosity mapping is a powerful technique for the identification of rare, autosomal recessive, disease-causing genes. The ease with which this category of disease gene can be identified has greatly increased through the availability of genome-wide SNP genotyping microarrays and subsequently of exome sequencing. Although these methods have simplified the generation of experimental data, its analysis, particularly when disparate data types must be integrated, remains time consuming. Moreover, the huge volume of sequence variant data generated from next generation sequencing experiments opens up the possibility of using these data instead of microarray genotype data to identify disease loci. To allow these two types of data to be used in an integrated fashion, we have developed AgileVCFMapper, a program that performs both the mapping of disease loci by SNP genotyping and the analysis of potentially deleterious variants using exome sequence variant data, in a single step. This method does not require microarray SNP genotype data, although analysis with a combination of microarray and exome genotype data enables more precise delineation of disease loci, due to superior marker density and distribution.

  15. High-throughput novel microsatellite marker of faba bean via next generation sequencing

    Directory of Open Access Journals (Sweden)

    Yang Tao

    2012-11-01

    Full Text Available Abstract Background Faba bean (Vicia faba L. is an important food legume crop, grown for human consumption globally including in China, Turkey, Egypt and Ethiopia. Although genetic gain has been made through conventional selection and breeding efforts, this could be substantially improved through the application of molecular methods. For this, a set of reliable molecular markers representative of the entire genome is required. Results A library with 125,559 putative SSR sequences was constructed and characterized for repeat type and length from a mixed genome of 247 spring and winter sown faba bean genotypes using 454 sequencing. A suit of 28,503 primer pair sequences were designed and 150 were randomly selected for validation. Of these, 94 produced reproducible amplicons that were polymorphic among 32 faba bean genotypes selected from diverse geographical locations. The number of alleles per locus ranged from 2 to 8, the expected heterozygocities ranged from 0.0000 to 1.0000, and the observed heterozygosities ranged from 0.0908 to 0.8410. The validation by UPGMA cluster analysis of 32 genotypes based on Nei's genetic distance, showed high quality and effectiveness of those novel SSR markers developed via next generation sequencing technology. Conclusions Large scale SSR marker development was successfully achieved using next generation sequencing of the V. faba genome. These novel markers are valuable for constructing genetic linkage maps, future QTL mapping, and marker-assisted trait selection in faba bean breeding efforts.

  16. Clinical Use of Next-Generation Sequencing in the Diagnosis of Wilson's Disease.

    Science.gov (United States)

    Németh, Dániel; Árvai, Kristóf; Horváth, Péter; Kósa, János Pál; Tobiás, Bálint; Balla, Bernadett; Folhoffer, Anikó; Krolopp, Anna; Lakatos, Péter András; Szalay, Ferenc

    2016-01-01

    Objective. Wilson's disease is a disorder of copper metabolism which is fatal without treatment. The great number of disease-causing ATP7B gene mutations and the variable clinical presentation of WD may cause a real diagnostic challenge. The emergence of next-generation sequencing provides a time-saving, cost-effective method for full sequencing of the whole ATP7B gene compared to the traditional Sanger sequencing. This is the first report on the clinical use of NGS to examine ATP7B gene. Materials and Methods. We used Ion Torrent Personal Genome Machine in four heterozygous patients for the identification of the other mutations and also in two patients with no known mutation. One patient with acute on chronic liver failure was a candidate for acute liver transplantation. The results were validated by Sanger sequencing. Results. In each case, the diagnosis of Wilson's disease was confirmed by identifying the mutations in both alleles within 48 hours. One novel mutation (p.Ala1270Ile) was found beyond the eight other known ones. The rapid detection of the mutations made possible the prompt diagnosis of WD in a patient with acute liver failure. Conclusions. According to our results we found next-generation sequencing a very useful, reliable, time-saving, and cost-effective method for diagnosing Wilson's disease in selected cases.

  17. The Quest for Rare Variants: Pooled Multiplexed Next Generation Sequencing in Plants

    Directory of Open Access Journals (Sweden)

    Fabio eMarroni

    2012-06-01

    Full Text Available Next generation sequencing (NGS instruments produce an unprecedented amount of sequence data at contained costs. This gives researchers the possibility of designing studies with adequate power to identify rare variants at a fraction of the economic and labor resources required by individual Sanger sequencing. As of today, only three research groups working in plant sciences have exploited this potentiality. They showed that pooled NGS can provide results in excellent agreement with those obtained by individual Sanger sequencing. Aim of this review is to convey to the reader the general ideas underlying the use of pooled NGS for the identification of rare variants. To facilitate a thorough understanding of the possibilities of the method we will explain in detail the variations in study design and discuss their advantages and disadvantages. We will show that information on allele frequency obtained by pooled next generation sequencing can be used to accurately compute basic population genetics indexes such as allele frequency, nucleotide diversity and Tajima’s D. Finally we will discuss applications and future perspectives of the multiplexed NGS approach.

  18. NGS catalog: A database of next generation sequencing studies in humans.

    Science.gov (United States)

    Xia, Junfeng; Wang, Qingguo; Jia, Peilin; Wang, Bing; Pao, William; Zhao, Zhongming

    2012-06-01

    Next generation sequencing (NGS) technologies have been rapidly applied in biomedical and biological research since its advent only a few years ago, and they are expected to advance at an unprecedented pace in the following years. To provide the research community with a comprehensive NGS resource, we have developed the database Next Generation Sequencing Catalog (NGS Catalog, http://bioinfo.mc.vanderbilt.edu/NGS/index.html), a continually updated database that collects, curates and manages available human NGS data obtained from published literature. NGS Catalog deposits publication information of NGS studies and their mutation characteristics (SNVs, small insertions/deletions, copy number variations, and structural variants), as well as mutated genes and gene fusions detected by NGS. Other functions include user data upload, NGS general analysis pipelines, and NGS software. NGS Catalog is particularly useful for investigators who are new to NGS but would like to take advantage of these powerful technologies for their own research. Finally, based on the data deposited in NGS Catalog, we summarized features and findings from whole exome sequencing, whole genome sequencing, and transcriptome sequencing studies for human diseases or traits.

  19. Clinical Use of Next-Generation Sequencing in the Diagnosis of Wilson’s Disease

    Directory of Open Access Journals (Sweden)

    Dániel Németh

    2016-01-01

    Full Text Available Objective. Wilson’s disease is a disorder of copper metabolism which is fatal without treatment. The great number of disease-causing ATP7B gene mutations and the variable clinical presentation of WD may cause a real diagnostic challenge. The emergence of next-generation sequencing provides a time-saving, cost-effective method for full sequencing of the whole ATP7B gene compared to the traditional Sanger sequencing. This is the first report on the clinical use of NGS to examine ATP7B gene. Materials and Methods. We used Ion Torrent Personal Genome Machine in four heterozygous patients for the identification of the other mutations and also in two patients with no known mutation. One patient with acute on chronic liver failure was a candidate for acute liver transplantation. The results were validated by Sanger sequencing. Results. In each case, the diagnosis of Wilson’s disease was confirmed by identifying the mutations in both alleles within 48 hours. One novel mutation (p.Ala1270Ile was found beyond the eight other known ones. The rapid detection of the mutations made possible the prompt diagnosis of WD in a patient with acute liver failure. Conclusions. According to our results we found next-generation sequencing a very useful, reliable, time-saving, and cost-effective method for diagnosing Wilson’s disease in selected cases.

  20. Next generation sequencing as a useful tool in the diagnostics of mosaicism in Alport syndrome.

    Science.gov (United States)

    Beicht, Sonja; Strobl-Wildemann, Gertrud; Rath, Sabine; Wachter, Oliver; Alberer, Martin; Kaminsky, Elke; Weber, Lutz T; Hinrichsen, Tanja; Klein, Hanns-Georg; Hoefele, Julia

    2013-09-10

    Alport syndrome (ATS) is a progressive hereditary nephropathy characterized by hematuria and/or proteinuria with structural defects of the glomerular basement membrane. It can be associated with extrarenal manifestations (high-tone sensorineural hearing loss and ocular abnormalities). Somatic mutations in COL4A5 (X-linked), COL4A3 and COL4A4 genes (both autosomal recessive and autosomal dominant) cause Alport syndrome. Somatic mosaicism in Alport patients is very rare. The reason for this may be due to the difficulty of detection. We report the case of a boy and his mother who presented with Alport syndrome. Mutational analysis showed the novel hemizygote pathogenic mutation c.2396-1G>A (IVS29-1G>A) at the splice acceptor site of the intron 29 exon 30 boundary of the COL4A5 gene in the boy. The mutation in the mother would not have been detected by Sanger sequencing without the knowledge of the mutational analysis result of her son. Further investigation of the mother using next generation sequencing showed somatic mosaicism and implied potential germ cell mosaicism. The mutation in the mother has most likely occurred during early embryogenesis. Analysis of tissue of different embryonic origin in the mother confirmed mosaicism in both mesoderm and ectoderm. Low grade mosaicism is very difficult to detect by Sanger sequencing. Next generation sequencing is increasingly used in the diagnostics and might improve the detection of mosaicism. In the case of definite clinical symptoms of ATS and missing detection of a mutation by Sanger sequencing, mutational analysis should be performed by next generation sequencing.

  1. IQSeq: integrated isoform quantification analysis based on next-generation sequencing.

    Directory of Open Access Journals (Sweden)

    Jiang Du

    Full Text Available With the recent advances in high-throughput RNA sequencing (RNA-Seq, biologists are able to measure transcription with unprecedented precision. One problem that can now be tackled is that of isoform quantification: here one tries to reconstruct the abundances of isoforms of a gene. We have developed a statistical solution for this problem, based on analyzing a set of RNA-Seq reads, and a practical implementation, available from archive.gersteinlab.org/proj/rnaseq/IQSeq, in a tool we call IQSeq (Isoform Quantification in next-generation Sequencing. Here, we present theoretical results which IQSeq is based on, and then use both simulated and real datasets to illustrate various applications of the tool. In order to measure the accuracy of an isoform-quantification result, one would try to estimate the average variance of the estimated isoform abundances for each gene (based on resampling the RNA-seq reads, and IQSeq has a particularly fast algorithm (based on the Fisher Information Matrix for calculating this, achieving a speedup of ~ 500 times compared to brute-force resampling. IQSeq also calculates an information theoretic measure of overall transcriptome complexity to describe isoform abundance for a whole experiment. IQSeq has many features that are particularly useful in RNA-Seq experimental design, allowing one to optimally model the integration of different sequencing technologies in a cost-effective way. In particular, the IQSeq formalism integrates the analysis of different sample (i.e. read sets generated from different technologies within the same statistical framework. It also supports a generalized statistical partial-sample-generation function to model the sequencing process. This allows one to have a modular, "plugin-able" read-generation function to support the particularities of the many evolving sequencing technologies.

  2. Application of next generation sequencing to human gene fusion detection: computational tools, features and perspectives.

    Science.gov (United States)

    Wang, Qingguo; Xia, Junfeng; Jia, Peilin; Pao, William; Zhao, Zhongming

    2013-07-01

    Gene fusions are important genomic events in human cancer because their fusion gene products can drive the development of cancer and thus are potential prognostic tools or therapeutic targets in anti-cancer treatment. Major advancements have been made in computational approaches for fusion gene discovery over the past 3 years due to improvements and widespread applications of high-throughput next generation sequencing (NGS) technologies. To identify fusions from NGS data, existing methods typically leverage the strengths of both sequencing technologies and computational strategies. In this article, we review the NGS and computational features of existing methods for fusion gene detection and suggest directions for future development.

  3. Aptaligner: automated software for aligning pseudorandom DNA X-aptamers from next-generation sequencing data.

    Science.gov (United States)

    Lu, Emily; Elizondo-Riojas, Miguel-Angel; Chang, Jeffrey T; Volk, David E

    2014-06-10

    Next-generation sequencing results from bead-based aptamer libraries have demonstrated that traditional DNA/RNA alignment software is insufficient. This is particularly true for X-aptamers containing specialty bases (W, X, Y, Z, ...) that are identified by special encoding. Thus, we sought an automated program that uses the inherent design scheme of bead-based X-aptamers to create a hypothetical reference library and Markov modeling techniques to provide improved alignments. Aptaligner provides this feature as well as length error and noise level cutoff features, is parallelized to run on multiple central processing units (cores), and sorts sequences from a single chip into projects and subprojects.

  4. Protection algorithm for a wind turbine generator based on positive- and negative-sequence fault components

    DEFF Research Database (Denmark)

    Zheng, Tai-Ying; Cha, Seung-Tae; Crossley, Peter A.;

    2011-01-01

    A protection relay for a wind turbine generator (WTG) based on positive- and negative-sequence fault components is proposed in the paper. The relay uses the magnitude of the positive-sequence component in the fault current to detect a fault on a parallel WTG, connected to the same power collection...... feeder, or a fault on an adjacent feeder; but for these faults, the relay remains stable and inoperative. A fault on the power collection feeder or a fault on the collection bus, both of which require an instantaneous tripping response, are distinguished from an inter-tie fault or a grid fault, which...

  5. Non-uniform phenotyping of D12S391 resolved by second generation sequencing

    DEFF Research Database (Denmark)

    Dalsgaard, S; Rockenbauer, E; Buchard, A;

    2014-01-01

    of the electropherograms suggested that the individuals were heterozygous with two alleles that differed in size by one nucleotide. This was confirmed by amplifying the samples with the PowerPlex(®) ESX 17 system. D12S391 is a complex STR with variable numbers of AGAT and AGAC repeats. Second generation sequencing...... revealed that separation of two alleles differing by one nucleotide in length was poor if the number of AGAT repeats in the short allele was higher than in the long allele. A total of 45 individuals with microvariants or off-ladder alleles in D12S391 were sequenced. Thirty different alleles were detected...

  6. The first insight into the tissue specific taxus transcriptome via Illumina second generation sequencing.

    Directory of Open Access Journals (Sweden)

    Da Cheng Hao

    Full Text Available BACKGROUND: Illumina second generation sequencing is now an efficient route for generating enormous sequence collections that represent expressed genes and quantitate expression level. Taxus is a world-wide endangered gymnosperm genus and forms an important anti-cancer medicinal resource, but the large and complex genomes of Taxus have hindered the development of genomic resources. The research of its tissue-specific transcriptome is absent. There is also no study concerning the association between the plant transcriptome and metabolome with respect to the plant tissue type. METHODOLOGY/PRINCIPAL FINDINGS: We performed the de novo assembly of Taxus mairei transcriptome using Illumina paired-end sequencing technology. In a single run, we produced 13,737,528 sequencing reads corresponding to 2.03 Gb total nucleotides. These reads were assembled into 36,493 unique sequences. Based on similarity search with known proteins, 23,515 Unigenes were identified to have the Blast hit with a cut-off E-value above 10⁻⁵. Furthermore, we investigated the transcriptome difference of three Taxus tissues using a tag-based digital gene expression system. We obtained a sequencing depth of over 3.15 million tags per sample and identified a large number of genes associated with tissue specific functions and taxane biosynthetic pathway. The expression of the taxane biosynthetic genes is significantly higher in the root than in the leaf and the stem, while high activity of taxane-producing pathway in the root was also revealed via metabolomic analyses. Moreover, many antisense transcripts and novel transcripts were found; clusters with similar differential expression patterns, enriched GO terms and enriched metabolic pathways with regard to the differentially expressed genes were revealed for the first time. CONCLUSIONS/SIGNIFICANCE: Our data provides the most comprehensive sequence resource available for Taxus study and will help define mechanisms of tissue

  7. Generating quantitative models describing the sequence specificity of biological processes with the stabilized matrix method

    Directory of Open Access Journals (Sweden)

    Sette Alessandro

    2005-05-01

    Full Text Available Abstract Background Many processes in molecular biology involve the recognition of short sequences of nucleic-or amino acids, such as the binding of immunogenic peptides to major histocompatibility complex (MHC molecules. From experimental data, a model of the sequence specificity of these processes can be constructed, such as a sequence motif, a scoring matrix or an artificial neural network. The purpose of these models is two-fold. First, they can provide a summary of experimental results, allowing for a deeper understanding of the mechanisms involved in sequence recognition. Second, such models can be used to predict the experimental outcome for yet untested sequences. In the past we reported the development of a method to generate such models called the Stabilized Matrix Method (SMM. This method has been successfully applied to predicting peptide binding to MHC molecules, peptide transport by the transporter associated with antigen presentation (TAP and proteasomal cleavage of protein sequences. Results Herein we report the implementation of the SMM algorithm as a publicly available software package. Specific features determining the type of problems the method is most appropriate for are discussed. Advantageous features of the package are: (1 the output generated is easy to interpret, (2 input and output are both quantitative, (3 specific computational strategies to handle experimental noise are built in, (4 the algorithm is designed to effectively handle bounded experimental data, (5 experimental data from randomized peptide libraries and conventional peptides can easily be combined, and (6 it is possible to incorporate pair interactions between positions of a sequence. Conclusion Making the SMM method publicly available enables bioinformaticians and experimental biologists to easily access it, to compare its performance to other prediction methods, and to extend it to other applications.

  8. Evaluation and Application of the Strand-Specific Protocol for Next-Generation Sequencing

    Directory of Open Access Journals (Sweden)

    Kuo-Wang Tsai

    2015-01-01

    Full Text Available Next-generation sequencing (NGS has become a powerful sequencing tool, applied in a wide range of biological studies. However, the traditional sample preparation protocol for NGS is non-strand-specific (NSS, leading to biased estimates of expression for transcripts overlapped at the antisense strand. Strand-specific (SS protocols have recently been developed. In this study, we prepared the same RNA sample by using the SS and NSS protocols, followed by sequencing with Illumina HiSeq platform. Using real-time quantitative PCR as a standard, we first proved that the SS protocol more precisely estimates gene expressions compared with the NSS protocol, particularly for those overlapped at the antisense strand. In addition, we also showed that the sequence reads from the SS protocol are comparable with those from conventional NSS protocols in many aspects. Finally, we also mapped a fraction of sequence reads back to the antisense strand of the known genes, originally without annotated genes located. Using sequence assembly and PCR validation, we succeeded in identifying and characterizing the novel antisense genes. Our results show that the SS protocol performs more accurately than the traditional NSS protocol and can be applied in future studies.

  9. Histoimmunogenetics Markup Language 1.0: Reporting next generation sequencing-based HLA and KIR genotyping.

    Science.gov (United States)

    Milius, Robert P; Heuer, Michael; Valiga, Daniel; Doroschak, Kathryn J; Kennedy, Caleb J; Bolon, Yung-Tsi; Schneider, Joel; Pollack, Jane; Kim, Hwa Ran; Cereb, Nezih; Hollenbach, Jill A; Mack, Steven J; Maiers, Martin

    2015-12-01

    We present an electronic format for exchanging data for HLA and KIR genotyping with extensions for next-generation sequencing (NGS). This format addresses NGS data exchange by refining the Histoimmunogenetics Markup Language (HML) to conform to the proposed Minimum Information for Reporting Immunogenomic NGS Genotyping (MIRING) reporting guidelines (miring.immunogenomics.org). Our refinements of HML include two major additions. First, NGS is supported by new XML structures to capture additional NGS data and metadata required to produce a genotyping result, including analysis-dependent (dynamic) and method-dependent (static) components. A full genotype, consensus sequence, and the surrounding metadata are included directly, while the raw sequence reads and platform documentation are externally referenced. Second, genotype ambiguity is fully represented by integrating Genotype List Strings, which use a hierarchical set of delimiters to represent allele and genotype ambiguity in a complete and accurate fashion. HML also continues to enable the transmission of legacy methods (e.g. site-specific oligonucleotide, sequence-specific priming, and Sequence Based Typing (SBT)), adding features such as allowing multiple group-specific sequencing primers, and fully leveraging techniques that combine multiple methods to obtain a single result, such as SBT integrated with NGS. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Coverage recommendation for genotyping analysis of highly heterologous species using next-generation sequencing technology

    Science.gov (United States)

    Song, Kai; Li, Li; Zhang, Guofan

    2016-01-01

    Next-generation sequencing (NGS) technology is being applied to an increasing number of non-model species and has been used as the primary approach for accurate genotyping in genetic and evolutionary studies. However, inferring genotypes from sequencing data is challenging, particularly for organisms with a high degree of heterozygosity. This is because genotype calls from sequencing data are often inaccurate due to low sequencing coverage, and if this is not accounted for, genotype uncertainty can lead to serious bias in downstream analyses, such as quantitative trait locus mapping and genome-wide association studies. Here, we used high-coverage reference data sets from Crassostrea gigas to simulate sequencing data with different coverage, and we evaluate the influence of genotype calling rate and accuracy as a function of coverage. Having initially identified the appropriate parameter settings for filtering to ensure genotype accuracy, we used two different single-nucleotide polymorphism (SNP) calling pipelines, single-sample and multi-sample. We found that a coverage of 15× was suitable for obtaining sufficient numbers of SNPs with high accuracy. Our work provides guidelines for the selection of sequence coverage when using NGS to investigate species with a high degree of heterozygosity and rapid decay of linkage disequilibrium. PMID:27760996

  11. Using Next Generation RAD Sequencing to Isolate Multispecies Microsatellites for Pilosocereus (Cactaceae).

    Science.gov (United States)

    Bonatelli, Isabel A S; Carstens, Bryan C; Moraes, Evandro M

    2015-01-01

    Microsatellite markers (also known as SSRs, Simple Sequence Repeats) are widely used in plant science and are among the most informative molecular markers for population genetic investigations, but the development of such markers presents substantial challenges. In this report, we discuss how next generation sequencing can replace the cloning, Sanger sequencing, identification of polymorphic loci, and testing cross-amplification that were previously required to develop microsatellites. We report the development of a large set of microsatellite markers for five species of the Neotropical cactus genus Pilosocereus using a restriction-site-associated DNA sequencing (RAD-seq) on a Roche 454 platform. We identified an average of 165 microsatellites per individual, with the absolute numbers across individuals proportional to the sequence reads obtained per individual. Frequency distribution of the repeat units was similar in the five species, with shorter motifs such as di- and trinucleotide being the most abundant repeats. In addition, we provide 72 microsatellites that could be potentially amplified in the sampled species and 22 polymorphic microsatellites validated in two populations of the species Pilosocereus machrisii. Although low coverage sequencing among individuals was observed for most of the loci, which we suggest to be more related to the nature of the microsatellite markers and the possible bias inserted by the restriction enzymes than to the genome size, our work demonstrates that an NGS approach is an efficient method to isolate multispecies microsatellites even in non-model organisms.

  12. Using Next Generation RAD Sequencing to Isolate Multispecies Microsatellites for Pilosocereus (Cactaceae.

    Directory of Open Access Journals (Sweden)

    Isabel A S Bonatelli

    Full Text Available Microsatellite markers (also known as SSRs, Simple Sequence Repeats are widely used in plant science and are among the most informative molecular markers for population genetic investigations, but the development of such markers presents substantial challenges. In this report, we discuss how next generation sequencing can replace the cloning, Sanger sequencing, identification of polymorphic loci, and testing cross-amplification that were previously required to develop microsatellites. We report the development of a large set of microsatellite markers for five species of the Neotropical cactus genus Pilosocereus using a restriction-site-associated DNA sequencing (RAD-seq on a Roche 454 platform. We identified an average of 165 microsatellites per individual, with the absolute numbers across individuals proportional to the sequence reads obtained per individual. Frequency distribution of the repeat units was similar in the five species, with shorter motifs such as di- and trinucleotide being the most abundant repeats. In addition, we provide 72 microsatellites that could be potentially amplified in the sampled species and 22 polymorphic microsatellites validated in two populations of the species Pilosocereus machrisii. Although low coverage sequencing among individuals was observed for most of the loci, which we suggest to be more related to the nature of the microsatellite markers and the possible bias inserted by the restriction enzymes than to the genome size, our work demonstrates that an NGS approach is an efficient method to isolate multispecies microsatellites even in non-model organisms.

  13. Calibrating convective-core overshooting with eclipsing binary systems. The case of low-mass main-sequence stars

    CERN Document Server

    Valle, G; Moroni, P G Prada; Degl'Innocenti, S

    2016-01-01

    In a robust statistical way, we quantify the uncertainty that affects the calibration of the overshooting efficiency parameter $\\beta$ that is owing to the uncertainty on the observational data in double-lined eclipsing binary systems. We also quantify the bias that is caused by the lack of constraints on the initial helium content and on the efficiencies of the superadiabatic convection and microscopic diffusion. We adopted a modified grid-based SCEPtER pipeline using as observational constraints the effective temperatures, [Fe/H], masses, and radii of the two stars. In a reference scenario of mild overshooting $\\beta = 0.2$ for the synthetic data, we found both large statistical uncertainties and biases on the estimated $\\beta$. For the first 80% of the MS evolution, $\\beta$ is biased and practically unconstrained in the whole explored range [0.0; 0.4]. In the last 5% of the MS the bias vanishes and the $1 \\sigma$ error is about 0.05. For synthetic data computed with $\\beta = 0.0$, the estimated $\\beta$ is ...

  14. A next generation semiconductor based sequencing approach for the identification of meat species in DNA mixtures.

    Science.gov (United States)

    Bertolini, Francesca; Ghionda, Marco Ciro; D'Alessandro, Enrico; Geraci, Claudia; Chiofalo, Vincenzo; Fontanesi, Luca

    2015-01-01

    The identification of the species of origin of meat and meat products is an important issue to prevent and detect frauds that might have economic, ethical and health implications. In this paper we evaluated the potential of the next generation semiconductor based sequencing technology (Ion Torrent Personal Genome Machine) for the identification of DNA from meat species (pig, horse, cattle, sheep, rabbit, chicken, turkey, pheasant, duck, goose and pigeon) as well as from human and rat in DNA mixtures through the sequencing of PCR products obtained from different couples of universal primers that amplify 12S and 16S rRNA mitochondrial DNA genes. Six libraries were produced including PCR products obtained separately from 13 species or from DNA mixtures containing DNA from all species or only avian or only mammalian species at equimolar concentration or at 1:10 or 1:50 ratios for pig and horse DNA. Sequencing obtained a total of 33,294,511 called nucleotides of which 29,109,688 with Q20 (87.43%) in a total of 215,944 reads. Different alignment algorithms were used to assign the species based on sequence data. Error rate calculated after confirmation of the obtained sequences by Sanger sequencing ranged from 0.0003 to 0.02 for the different species. Correlation about the number of reads per species between different libraries was high for mammalian species (0.97) and lower for avian species (0.70). PCR competition limited the efficiency of amplification and sequencing for avian species for some primer pairs. Detection of low level of pig and horse DNA was possible with reads obtained from different primer pairs. The sequencing of the products obtained from different universal PCR primers could be a useful strategy to overcome potential problems of amplification. Based on these results, the Ion Torrent technology can be applied for the identification of meat species in DNA mixtures.

  15. Stepwise threshold clustering: a new method for genotyping MHC loci using next-generation sequencing technology.

    Directory of Open Access Journals (Sweden)

    William E Stutz

    Full Text Available Genes of the vertebrate major histocompatibility complex (MHC are of great interest to biologists because of their important role in immunity and disease, and their extremely high levels of genetic diversity. Next generation sequencing (NGS technologies are quickly becoming the method of choice for high-throughput genotyping of multi-locus templates like MHC in non-model organisms. Previous approaches to genotyping MHC genes using NGS technologies suffer from two problems:1 a "gray zone" where low frequency alleles and high frequency artifacts can be difficult to disentangle and 2 a similar sequence problem, where very similar alleles can be difficult to distinguish as two distinct alleles. Here were present a new method for genotyping MHC loci--Stepwise Threshold Clustering (STC--that addresses these problems by taking full advantage of the increase in sequence data provided by NGS technologies. Unlike previous approaches for genotyping MHC with NGS data that attempt to classify individual sequences as alleles or artifacts, STC uses a quasi-Dirichlet clustering algorithm to cluster similar sequences at increasing levels of sequence similarity. By applying frequency and similarity based criteria to clusters rather than individual sequences, STC is able to successfully identify clusters of sequences that correspond to individual or similar alleles present in the genomes of individual samples. Furthermore, STC does not require duplicate runs of all samples, increasing the number of samples that can be genotyped in a given project. We show how the STC method works using a single sample library. We then apply STC to 295 threespine stickleback (Gasterosteus aculeatus samples from four populations and show that neighboring populations differ significantly in MHC allele pools. We show that STC is a reliable, accurate, efficient, and flexible method for genotyping MHC that will be of use to biologists interested in a variety of downstream applications.

  16. A next generation semiconductor based sequencing approach for the identification of meat species in DNA mixtures.

    Directory of Open Access Journals (Sweden)

    Francesca Bertolini

    Full Text Available The identification of the species of origin of meat and meat products is an important issue to prevent and detect frauds that might have economic, ethical and health implications. In this paper we evaluated the potential of the next generation semiconductor based sequencing technology (Ion Torrent Personal Genome Machine for the identification of DNA from meat species (pig, horse, cattle, sheep, rabbit, chicken, turkey, pheasant, duck, goose and pigeon as well as from human and rat in DNA mixtures through the sequencing of PCR products obtained from different couples of universal primers that amplify 12S and 16S rRNA mitochondrial DNA genes. Six libraries were produced including PCR products obtained separately from 13 species or from DNA mixtures containing DNA from all species or only avian or only mammalian species at equimolar concentration or at 1:10 or 1:50 ratios for pig and horse DNA. Sequencing obtained a total of 33,294,511 called nucleotides of which 29,109,688 with Q20 (87.43% in a total of 215,944 reads. Different alignment algorithms were used to assign the species based on sequence data. Error rate calculated after confirmation of the obtained sequences by Sanger sequencing ranged from 0.0003 to 0.02 for the different species. Correlation about the number of reads per species between different libraries was high for mammalian species (0.97 and lower for avian species (0.70. PCR competition limited the efficiency of amplification and sequencing for avian species for some primer pairs. Detection of low level of pig and horse DNA was possible with reads obtained from different primer pairs. The sequencing of the products obtained from different universal PCR primers could be a useful strategy to overcome potential problems of amplification. Based on these results, the Ion Torrent technology can be applied for the identification of meat species in DNA mixtures.

  17. SPARSE SEQUENCE CONSTRUCTION OF LDPC CODES

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    This letter proposes a novel and simple construction of regular Low-Density Parity-Check (LDPC) codes using sparse binary sequences. It utilizes the cyclic cross correlation function of sparse sequences to generate codes with girth8. The new codes perform well using the sumproduct decoding. Low encodingcomplexity can also be achieved due to the inherent quasi-cyclic structure of the codes.

  18. Solving the molecular diagnostic testing conundrum for Mendelian disorders in the era of next-generation sequencing: single-gene, gene panel, or exome/genome sequencing.

    Science.gov (United States)

    Xue, Yuan; Ankala, Arunkanth; Wilcox, William R; Hegde, Madhuri R

    2015-06-01

    Next-generation sequencing is changing the paradigm of clinical genetic testing. Today there are numerous molecular tests available, including single-gene tests, gene panels, and exome sequencing or genome sequencing. As a result, ordering physicians face the conundrum of selecting the best diagnostic tool for their patients with genetic conditions. Single-gene testing is often most appropriate for conditions with distinctive clinical features and minimal locus heterogeneity. Next-generation sequencing-based gene panel testing, which can be complemented with array comparative genomic hybridization and other ancillary methods, provides a comprehensive and feasible approach for heterogeneous disorders. Exome sequencing and genome sequencing have the advantage of being unbiased regarding what set of genes is analyzed, enabling parallel interrogation of most of the genes in the human genome. However, current limitations of next-generation sequencing technology and our variant interpretation capabilities caution us against offering exome sequencing or genome sequencing as either stand-alone or first-choice diagnostic approaches. A growing interest in personalized medicine calls for the application of genome sequencing in clinical diagnostics, but major challenges must be addressed before its full potential can be realized. Here, we propose a testing algorithm to help clinicians opt for the most appropriate molecular diagnostic tool for each scenario.

  19. Texture and mechanical properties of tape substrates from binary and ternary copper alloys for second-generation high-temperature superconductors

    Science.gov (United States)

    Khlebnikova, Yu. V.; Rodionov, D. P.; Gervas'eva, I. V.; Suaridze, T. R.; Egorova, L. Yu.; Akshentsev, Yu. N.; Kazantsev, V. A.

    2015-01-01

    The process of texture formation in tapes made of a number of binary and ternary copper alloys upon cold rolling to degrees of deformation of 98.6-99% and subsequent recrystallization annealing has been studied. The possibility of designing multicomponent alloys based on the binary Cu-30% Ni alloy additionally alloyed with elements that strengthen the fcc matrix, such as iron or chromium, has been shown. The opportunity of obtaining a perfect cube texture in a thin tape made of binary and ternary copper alloys opens prospects for their use as substrates in the technology of second-generation HTSC cables. Optimum regimes of annealing have been determined, which make it possible to obtain in the Cu- M and Cu-(30-40)Ni- M ( M = Fe, Cr, Mn) alloys a perfect biaxial texture with the fraction of cube grains {001} on the surface of the tape more than 94%. The estimation of the mechanical properties of the textured tapes of the investigated alloys demonstrates a yield strength that is 2.5-4.5 times greater than that in the textured tape of pure copper.

  20. Substitute CT generation from a single ultra short time echo MRI sequence: preliminary study

    Science.gov (United States)

    Ghose, Soumya; Dowling, Jason A.; Rai, Robba; Liney, Gary P.

    2017-04-01

    In MR guided radiation therapy planning both MR and CT images for a patient are acquired and co-registered to obtain a tissue specific HU map. Generation of the HU map directly from the MRI would eliminate the CT acquisition and may improve radiation therapy planning. In this preliminary study of substitute CT (sCT) generation, two porcine leg phantoms were scanned using a 3D ultrashort echo time (PETRA) sequence and co-registered to corresponding CT images to build tissue specific regression models. The model was created from one co-registered CT-PETRA pair to generate the sCT for the other PETRA image. An expectation maximization based clustering was performed on the co-registered PETRA image to identify the soft tissues, dense bone and air class membership probabilities. A tissue specific non linear regression model was built from one registered CT-PETRA pair dataset to predict the sCT of the second PETRA image in a two-fold cross validation schema. A complete substitute CT is generated in 3 min. The mean absolute HU error for air was 0.3 HU, bone was 95 HU, fat was 30 HU and for muscle it was 10 HU. The mean surface reconstruction error for the bone was 1.3 mm. The PETRA sequence enabled a low mean absolute surface distance for the bone and a low HU error for other classes. The sCT generated from a single PETRA sequence shows promise for the generation of fast sCT for MRI based radiation therapy planning.

  1. Microsatellites for next-generation ecologists: a post-sequencing bioinformatics pipeline.

    Science.gov (United States)

    Fernandez-Silva, Iria; Whitney, Jonathan; Wainwright, Benjamin; Andrews, Kimberly R; Ylitalo-Ward, Heather; Bowen, Brian W; Toonen, Robert J; Goetze, Erica; Karl, Stephen A

    2013-01-01

    Microsatellites are the markers of choice for a variety of population genetic studies. The recent advent of next-generation pyrosequencing has drastically accelerated microsatellite locus discovery by providing a greater amount of DNA sequencing reads at lower costs compared to other techniques. However, laboratory testing of PCR primers targeting potential microsatellite markers remains time consuming and costly. Here we show how to reduce this workload by screening microsatellite loci via bioinformatic analyses prior to primer design. Our method emphasizes the importance of sequence quality, and we avoid loci associated with repetitive elements by screening with repetitive sequence databases available for a growing number of taxa. Testing with the Yellowstripe Goatfish Mulloidichthys flavolineatus and the marine planktonic copepod Pleuromamma xiphias we show higher success rate of primers selected by our pipeline in comparison to previous in silico microsatellite detection methodologies. Following the same pipeline, we discover and select microsatellite loci in nine additional species including fishes, sea stars, copepods and octopuses.

  2. The Application of Next Generation Sequencing Technology on Noninvasive Prenatal Test

    DEFF Research Database (Denmark)

    Jiang, Hui

    of effective treatment. The rapid development of next generation sequencing technology boosts the discovery of new causative gene for these rare diseases, as well as the genetic diagnosis in clinic practice. Carrier screening, prenatal diagnosis and newborn screening are wildly used in the world to prevent...... an invasive process, which might lead to maternal anxiety, or even miscarriage. Therefore, developing an effective approach to perform noninvasive prenatal test (NIPT) for rare diseases is the key challenge to prevent birth defect in the future. The discovery of cell-­free fetal DNA, coupling with next......, and maternal plasma. In order to obtain accurate result, we combined the haplotype information from the parents with maternal plasma deep sequencing data to recover the fetal genotype. Our study demonstrated that the sequencing-based new approach could be used to detect rare diseases, including chromosomal...

  3. Cracking the Code of Human Diseases Using Next-Generation Sequencing: Applications, Challenges, and Perspectives

    Directory of Open Access Journals (Sweden)

    Vincenza Precone

    2015-01-01

    Full Text Available Next-generation sequencing (NGS technologies have greatly impacted on every field of molecular research mainly because they reduce costs and increase throughput of DNA sequencing. These features, together with the technology’s flexibility, have opened the way to a variety of applications including the study of the molecular basis of human diseases. Several analytical approaches have been developed to selectively enrich regions of interest from the whole genome in order to identify germinal and/or somatic sequence variants and to study DNA methylation. These approaches are now widely used in research, and they are already being used in routine molecular diagnostics. However, some issues are still controversial, namely, standardization of methods, data analysis and storage, and ethical aspects. Besides providing an overview of the NGS-based approaches most frequently used to study the molecular basis of human diseases at DNA level, we discuss the principal challenges and applications of NGS in the field of human genomics.

  4. Next-generation sequencing and genetic diagnosis of Charcot-Marie-Tooth disease

    Directory of Open Access Journals (Sweden)

    Ashok Verma

    2014-01-01

    Full Text Available Over 70 different Charcot-Marie-Tooth disease (CMT–associated genes have now been discovered and their number is growing. Conventional genetic testing for all CMT genes is cumbersome, expensive, and impractical in an individual patient. Next-generation sequencing (NGS technology allows cost-effective sequencing of large scale DNA, even entire exome (coding sequences or whole genome and thus, NGS platform can be employed to effectively target a large number or all CMT-related genes for accurate diagnosis. This overview discusses how NGS can be strategically used for genetic diagnosis in patients with CMT or unexplained neuropathy. A comment is made to combine simple clinical and electrophysiological algorithm to assign patients to major CMT subtypes and then employ NGS to screen for all known mutations in the subtype-specific CMT gene panel.

  5. Next generation sequencing in the clinical domain: clinical advantages, practical, and ethical challenges.

    Science.gov (United States)

    Thompson, Rose; Drew, Cheney J G; Thomas, Rhys H

    2012-01-01

    There has been an academic "gold rush" with researchers mining the deep seams of whole-exome and whole-genome sequencing since 2008. Although undoubtedly a major advance initially for identifying new disease-associated genes for rare monogenetic disorders--more recently, common and complex conditions have been successfully studied using these techniques. With great power comes great responsibility, however, and we must not forget that next generation sequencing produces unique ethical conundrums and validation challenges. We review the progression of published papers using whole-exome sequencing from a clinical and technical viewpoint before then reflecting on the key arguments that need to be fully understood before these tools can become a routine part of clinical practice and we ask what may be the role for the biomedical scientists?

  6. Different next generation sequencing platforms produce different microbial profiles and diversity in cystic fibrosis sputum.

    Science.gov (United States)

    Hahn, Andrea; Sanyal, Amit; Perez, Geovanny F; Colberg-Poley, Anamaris M; Campos, Joseph; Rose, Mary C; Pérez-Losada, Marcos

    2016-11-01

    Cystic fibrosis (CF) is an autosomal recessive disease characterized by recurrent lung infections. Studies of the lung microbiome have shown an association between decreasing diversity and progressive disease. 454 pyrosequencing has frequently been used to study the lung microbiome in CF, but will no longer be supported. We sought to identify the benefits and drawbacks of using two state-of-the-art next generation sequencing (NGS) platforms, MiSeq and PacBio RSII, to characterize the CF lung microbiome. Each has its advantages and limitations. Twelve samples of extracted bacterial DNA were sequenced on both MiSeq and PacBio NGS platforms. DNA was amplified for the V4 region of the 16S rRNA gene and libraries were sequenced on the MiSeq sequencing platform, while the full 16S rRNA gene was sequenced on the PacBio RSII sequencing platform. Raw FASTQ files generated by the MiSeq and PacBio platforms were processed in mothur v1.35.1. There was extreme discordance in alpha-diversity of the CF lung microbiome when using the two platforms. Because of its depth of coverage, sequencing of the 16S rRNA V4 gene region using MiSeq allowed for the observation of many more operational taxonomic units (OTUs) and higher Chao1 and Shannon indices than the PacBio RSII. Interestingly, several patients in our cohort had Escherichia, an unusual pathogen in CF. Also, likely because of its coverage of the complete 16S rRNA gene, only PacBio RSII was able to identify Burkholderia, an important CF pathogen. When comparing microbiome diversity in clinical samples from CF patients using 16S sequences, MiSeq and PacBio NGS platforms may generate different results in microbial community composition and structure. It may be necessary to use different platforms when trying to correctly identify dominant pathogens versus measuring alpha-diversity estimates, and it would be important to use the same platform for comparisons to minimize errors in interpretation. Copyright © 2016 Elsevier B.V. All

  7. Next generation sequencing for molecular diagnosis of neurological disorders using ataxias as a model.

    Science.gov (United States)

    Németh, Andrea H; Kwasniewska, Alexandra C; Lise, Stefano; Parolin Schnekenberg, Ricardo; Becker, Esther B E; Bera, Katarzyna D; Shanks, Morag E; Gregory, Lorna; Buck, David; Zameel Cader, M; Talbot, Kevin; de Silva, Rajith; Fletcher, Nicholas; Hastings, Rob; Jayawant, Sandeep; Morrison, Patrick J; Worth, Paul; Taylor, Malcolm; Tolmie, John; O'Regan, Mary; Valentine, Ruth; Packham, Emily; Evans, Julie; Seller, Anneke; Ragoussis, Jiannis

    2013-10-01

    Many neurological conditions are caused by immensely heterogeneous gene mutations. The diagnostic process is often long and complex with most patients undergoing multiple invasive and costly investigations without ever reaching a conclusive molecular diagnosis. The advent of massively parallel, next-generation sequencing promises to revolutionize genetic testing and shorten the 'diagnostic odyssey' for many of these patients. We performed a pilot study using heterogeneous ataxias as a model neurogenetic disorder to assess the introduction of next-generation sequencing into clinical practice. We captured 58 known human ataxia genes followed by Illumina Next-Generation Sequencing in 50 highly heterogeneous patients with ataxia who had been extensively investigated and were refractory to diagnosis. All cases had been tested for spinocerebellar ataxia 1-3, 6, 7 and Friedrich's ataxia and had multiple other biochemical, genetic and invasive tests. In those cases where we identified the genetic mutation, we determined the time to diagnosis. Pathogenicity was assessed using a bioinformatics pipeline and novel variants were validated using functional experiments. The overall detection rate in our heterogeneous cohort was 18% and varied from 8.3% in those with an adult onset progressive disorder to 40% in those with a childhood or adolescent onset progressive disorder. The highest detection rate was in those with an adolescent onset and a family history (75%). The majority of cases with detectable mutations had a childhood onset but most are now adults, reflecting the long delay in diagnosis. The delays were primarily related to lack of easily available clinical testing, but other factors included the presence of atypical phenotypes and the use of indirect testing. In the cases where we made an eventual diagnosis, the delay was 3-35 years (mean 18.1 years). Alignment and coverage metrics indicated that the capture and sequencing was highly efficient and the consumable cost

  8. Suppression Subtractive Hybridization Versus Next-Generation Sequencing in Plant Genetic Engineering: Challenges and Perspectives.

    Science.gov (United States)

    Sahebi, Mahbod; Hanafi, Mohamed M; Azizi, Parisa; Hakim, Abdul; Ashkani, Sadegh; Abiri, Rambod

    2015-10-01

    Suppression subtractive hybridization (SSH) is an effective method to identify different genes with different expression levels involved in a variety of biological processes. This method has often been used to study molecular mechanisms of plants in complex relationships with different pathogens and a variety of biotic stresses. Compared to other techniques used in gene expression profiling, SSH needs relatively smaller amounts of the initial materials, with lower costs, and fewer false positives present within the results. Extraction of total RNA from plant species rich in phenolic compounds, carbohydrates, and polysaccharides that easily bind to nucleic acids through cellular mechanisms is difficult and needs to be considered. Remarkable advancement has been achieved in the next-generation sequencing (NGS) field. As a result of progress within fields related to molecular chemistry and biology as well as specialized engineering, parallelization in the sequencing reaction has exceptionally enhanced the overall read number of generated sequences per run. Currently available sequencing platforms support an earlier unparalleled view directly into complex mixes associated with RNA in addition to DNA samples. NGS technology has demonstrated the ability to sequence DNA with remarkable swiftness, therefore allowing previously unthinkable scientific accomplishments along with novel biological purposes. However, the massive amounts of data generated by NGS impose a substantial challenge with regard to data safe-keeping and analysis. This review examines some simple but vital points involved in preparing the initial material for SSH and introduces this method as well as its associated applications to detect different novel genes from different plant species. This review evaluates general concepts, basic applications, plus the probable results of NGS technology in genomics, with unique mention of feasible potential tools as well as bioinformatics.

  9. Genome-wide characterization of pancreatic adenocarcinoma patients using next generation sequencing.

    Directory of Open Access Journals (Sweden)

    Winnie S Liang

    Full Text Available Pancreatic adenocarcinoma (PAC is among the most lethal malignancies. While research has implicated multiple genes in disease pathogenesis, identification of therapeutic leads has been difficult and the majority of currently available therapies provide only marginal benefit. To address this issue, our goal was to genomically characterize individual PAC patients to understand the range of aberrations that are occurring in each tumor. Because our understanding of PAC tumorigenesis is limited, evaluation of separate cases may reveal aberrations, that are less common but may provide relevant information on the disease, or that may represent viable therapeutic targets for the patient. We used next generation sequencing to assess global somatic events across 3 PAC patients to characterize each patient and to identify potential targets. This study is the first to report whole genome sequencing (WGS findings in paired tumor/normal samples collected from 3 separate PAC patients. We generated on average 132 billion mappable bases across all patients using WGS, and identified 142 somatic coding events including point mutations, insertion/deletions, and chromosomal copy number variants. We did not identify any significant somatic translocation events. We also performed RNA sequencing on 2 of these patients' tumors for which tumor RNA was available to evaluate expression changes that may be associated with somatic events, and generated over 100 million mapped reads for each patient. We further performed pathway analysis of all sequencing data to identify processes that may be the most heavily impacted from somatic and expression alterations. As expected, the KRAS signaling pathway was the most heavily impacted pathway (P<0.05, along with tumor-stroma interactions and tumor suppressive pathways. While sequencing of more patients is needed, the high resolution genomic and transcriptomic information we have acquired here provides valuable information on the

  10. Next-generation sequencing for diagnosis of rare diseases in the neonatal intensive care unit

    Science.gov (United States)

    Daoud, Hussein; Luco, Stephanie M.; Li, Rui; Bareke, Eric; Beaulieu, Chandree; Jarinova, Olga; Carson, Nancy; Nikkel, Sarah M.; Graham, Gail E.; Richer, Julie; Armour, Christine; Bulman, Dennis E.; Chakraborty, Pranesh; Geraghty, Michael; Lines, Matthew A.; Lacaze-Masmonteil, Thierry; Majewski, Jacek; Boycott, Kym M.; Dyment, David A.

    2016-01-01

    Background: Rare diseases often present in the first days and weeks of life and may require complex management in the setting of a neonatal intensive care unit (NICU). Exhaustive consultations and traditional genetic or metabolic investigations are costly and often fail to arrive at a final diagnosis when no recognizable syndrome is suspected. For this pilot project, we assessed the feasibility of next-generation sequencing as a tool to improve the diagnosis of rare diseases in newborns in the NICU. Methods: We retrospectively identified and prospectively recruited newborns and infants admitted to the NICU of the Children’s Hospital of Eastern Ontario and the Ottawa Hospital, General Campus, who had been referred to the medical genetics or metabolics inpatient consult service and had features suggesting an underlying genetic or metabolic condition. DNA from the newborns and parents was enriched for a panel of clinically relevant genes and sequenced on a MiSeq sequencing platform (Illumina Inc.). The data were interpreted with a standard informatics pipeline and reported to care providers, who assessed the importance of genotype–phenotype correlations. Results: Of 20 newborns studied, 8 received a diagnosis on the basis of next-generation sequencing (diagnostic rate 40%). The diagnoses were renal tubular dysgenesis, SCN1A-related encephalopathy syndrome, myotubular myopathy, FTO deficiency syndrome, cranioectodermal dysplasia, congenital myasthenic syndrome, autosomal dominant intellectual disability syndrome type 7 and Denys–Drash syndrome. Interpretation: This pilot study highlighted the potential of next-generation sequencing to deliver molecular diagnoses rapidly with a high success rate. With broader use, this approach has the potential to alter health care delivery in the NICU. PMID:27241786

  11. Targeted recovery of novel phylogenetic diversity from next-generation sequence data.

    Science.gov (United States)

    Lynch, Michael D J; Bartram, Andrea K; Neufeld, Josh D

    2012-11-01

    Next-generation sequencing technologies have led to recognition of a so-called 'rare biosphere'. These microbial operational taxonomic units (OTUs) are defined by low relative abundance and may be specifically adapted to maintaining low population sizes. We hypothesized that mining of low-abundance next-generation 16S ribosomal RNA (rRNA) gene data would lead to the discovery of novel phylogenetic diversity, reflecting microorganisms not yet discovered by previous sampling efforts. Here, we test this hypothesis by combining molecular and bioinformatic approaches for targeted retrieval of phylogenetic novelty within rare biosphere OTUs. We combined BLASTN network analysis, phylogenetics and targeted primer design to amplify 16S rRNA gene sequences from unique potential bacterial lineages, comprising part of the rare biosphere from a multi-million sequence data set from an Arctic tundra soil sample. Demonstrating the feasibility of the protocol developed here, three of seven recovered phylogenetic lineages represented extremely divergent taxonomic entities. These divergent target sequences correspond to (a) a previously unknown lineage within the BRC1 candidate phylum, (b) a sister group to the early diverging and currently recognized monospecific Cyanobacteria Gloeobacter, a genus containing multiple plesiomorphic traits and (c) a highly divergent lineage phylogenetically resolved within mitochondria. A comparison to twelve next-generation data sets from additional soils suggested persistent low-abundance distributions of these novel 16S rRNA genes. The results demonstrate this sequence analysis and retrieval pipeline as applicable for exploring underrepresented phylogenetic novelty and recovering taxa that may represent significant steps in bacterial evolution.

  12. Genotyping of simple sequence repeats--factors implicated in shadow band generation revisited.

    Science.gov (United States)

    Olejniczak, Marta; Krzyzosiak, Wlodzimierz J

    2006-10-01

    PCR amplification of microsatellite sequences generates, besides the main product corresponding to allele size, also additional, undesired products usually shorter by multiples of the repeated unit. These extra products known as shadow bands or stutter products may complicate genotyping. The mechanism by which these artifacts are formed is not well understood and so no effective remedy has been found to cope with these spurious products. In this study, using the DNA templates containing the CAG/CTG repeats flanked by gene-specific sequences and universal priming sites, we analyzed the effects of many PCR variables on the shadow band generation. The most important result was that at the decreased temperature of the denaturation step during PCR cycling the shadow bands were either not formed or were strongly suppressed. Several possible sources of this effect are discussed.

  13. Perspectives of DNA microarray and next-generation DNA sequencing technologies

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    DNA microarray and next-generation DNA sequencing technologies are important tools for high-throughput genome research,in revealing both the structural and functional characteristics of genomes.In the past decade the DNA microarray technologies have been widely applied in the studies of functional genomics,systems biology and pharmacogenomics.The next-generation DNA sequencing method was first introduced by the 454 Company in 2003,immediately followed by the establishment of the Solexa and Solid techniques by other biotech companies.Though it has not been long since the first emergence of this technology,with the fast and impressive improvement,the application of this technology has extended to almost all fields of genomics research,as a rival challenging the existing DNA microarray technology.This paper briefly reviews the working principles of these two technologies as well as their application and perspectives in genome research.

  14. Image encryption combining multiple generating sequences controlled fractional DCT with dependent scrambling and diffusion

    Science.gov (United States)

    Liang, Yaru; Liu, Guoping; Zhou, Nanrun; Wu, Jianhua

    2015-02-01

    Based on the fractional discrete cosine transform with multiple generating sequences (MGSFrDCT) and the dependent scrambling and diffusion (DSD), an image encryption algorithm is proposed, in which the multiple-generating sequences greatly enlarge the key space of the encryption system. The real-valued output of MGSFrDCT is beneficial to storage, display and transmission of the cipher-text. During the stage of confusion and diffusion, the locations and values of all MGSFrDCT transformed coefficients change due to DSD, and the initial values and fractional orders of encryption system depend not only on the cipher keys but also on the plain-image due to introduction of a disturbance factor, which allows the encryption system to resist the known-plaintext and chosen-plaintext attacks. Experimental results demonstrate that the proposed encryption algorithm is feasible, effective and secure and able to resist common classical attacks.

  15. Next Generation Sequencing As an Aid to Diagnosis and Treatment of an Unusual Pediatric Brain Cancer

    Directory of Open Access Journals (Sweden)

    John Glod

    2014-07-01

    Full Text Available Classification of pediatric brain tumors with unusual histologic and clinical features may be a diagnostic challenge to the pathologist. We present a case of a 12-year-old girl with a primary intracranial tumor. The tumor classification was not certain initially, and the site of origin and clinical behavior were unusual. Genomic characterization of the tumor using a Clinical Laboratory Improvement Amendment (CLIA-certified next-generation sequencing assay assisted in the diagnosis and translated into patient benefit, albeit transient. Our case argues that next generation sequencing may play a role in the pathological classification of pediatric brain cancers and guiding targeted therapy, supporting additional studies of genetically targeted therapeutics.

  16. Achieving high throughput sequencing of a cDNA library utilizing an alternative protocol for the bench top next-generation sequencing system.

    Science.gov (United States)

    Wan, Minxi; Faruq, Junaid; Rosenberg, Julian N; Xia, Jinlan; Oyler, George A; Betenbaugh, Michael J

    2013-02-15

    The development of next-generation sequencing (NGS) technologies has provided novel tools for genome analysis and expression profiling. A high throughput cDNA sequencing method using a bench top next-generation sequencing system, GS Junior, is now available. Here, we used an alternative protocol to the standard method for generating the cDNA library. This protocol can decrease the number of processing steps to manipulate RNA when constructing a cDNA library from an RNA sample, and does not require mRNA isolation from total RNA. Thus it can decrease the risk of RNA degradation and the cost for preparing a cDNA library. Also, the efficiency of sequencing data obtained with this approach is comparable to the standard method as verified by sequencing characteristics and expression levels of the reference gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

  17. Next-generation sequencing as a powerful motor for advances in the biological and environmental sciences.

    Science.gov (United States)

    Faure, Denis; Joly, Dominique

    2015-04-01

    Next-generation sequencing (NGS) provides unprecedented insight into (meta)genomes, (meta)transcriptomes (cDNA) and (meta)barcodes of individuals, populations and communities of Archaea, Bacteria and Eukarya, as well as viruses. This special issue combines reviews and original papers reporting technical and scientific advances in genomics and transcriptomics of non-model species, as well as quantification and functional analyses of biodiversity using NGS technologies of the second and third generations. In addition, certain papers also exemplify the transition from Sanger to NGS barcodes in molecular taxonomy.

  18. INTEGRATED APPROACH TO GENERATION OF PRECEDENCE RELATIONS AND PRECEDENCE GRAPHS FOR ASSEMBLY SEQUENCE PLANNING

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    An integrated approach to generation of precedence relations and precedence graphs for assembly sequence planning is presented, which contains more assembly flexibility. The approach involves two stages. Based on the assembly model, the components in the assembly can be divided into partially constrained components and completely constrained components in the first stage, and then geometric precedence relation for every component is generated automatically. According to the result of the first stage, the second stage determines and constructs all precedence graphs. The algorithms of these two stages proposed are verified by two assembly examples.

  19. rMotifGen: random motif generator for DNA and protein sequences

    Directory of Open Access Journals (Sweden)

    Hardin C Timothy

    2007-08-01

    Full Text Available Abstract Background Detection of short, subtle conserved motif regions within a set of related DNA or amino acid sequences can lead to discoveries about important regulatory domains such as transcription factor and DNA binding sites as well as conserved protein domains. In order to help assess motif detection algorithms on motifs with varying properties and levels of conservation, we have developed a computational tool, rMotifGen, with the sole purpose of generating a number of random DNA or protein sequences containing short sequence motifs. Each motif consensus can be user-defined, randomly generated, or created from a position-specific scoring matrix (PSSM. Insertions and mutations within these motifs are created according to user-defined parameters and substitution matrices. The resulting sequences can be helpful in mutational simulations and in testing the limits of motif detection algorithms. Results Two implementations of rMotifGen have been created, one providing a graphical user interface (GUI for random motif construction, and the other serving as a command line interface. The second implementation has the added advantages of platform independence and being able to be called in a batch mode. rMotifGen was used to construct sample sets of sequences containing DNA motifs and amino acid motifs that were then tested against the Gibbs sampler and MEME packages. Conclusion rMotifGen provides an efficient and convenient method for creating random DNA or amino acid sequences with a variable number of motifs, where the instance of each motif can be incorporated using a position-specific scoring matrix (PSSM or by creating an instance mutated from its corresponding consensus using an evolutionary model based on substitution matrices. rMotifGen is freely available at: http://bioinformatics.louisville.edu/brg/rMotifGen/.

  20. Generation and analysis of expressed sequence tags (ESTs) for marker development in yam (Dioscorea alata L.).

    Science.gov (United States)

    Narina, Satya S; Buyyarapu, Ramesh; Kottapalli, Kameswara Rao; Sartie, Alieu M; Ali, Mohamed I; Robert, Asiedu; Hodeba, Mignouna J D; Sayre, Brian L; Scheffler, Brian E

    2011-02-09

    Anthracnose (Colletotrichum gloeosporioides) is a major limiting factor in the production of yam (Dioscorea spp.) worldwide. Availability of high quality sequence information is necessary for designing molecular markers associated with resistance. However, very limited sequence information pertaining to yam is available at public genome databases. Therefore, this collaborative project was developed for genetic improvement and germplasm characterization of yams using molecular markers. The current investigation is focused on studying gene expression, by large scale generation of ESTs, from one susceptible (TDa 95-0310) and two resistant yam genotypes (TDa 87-01091, TDa 95-0328) challenged with the fungus. Total RNA was isolated from young leaves of resistant and susceptible genotypes and cDNA libraries were sequenced using Roche 454 technology. A total of 44,757 EST sequences were generated from the cDNA libraries of the resistant and susceptible genotypes. Greater than 56% of ESTs were annotated using MapMan Mercator tool and Blast2GO search tools. Gene annotations were used to characterize the transcriptome in yam and also perform a differential gene expression analysis between the resistant and susceptible EST datasets. Mining for SSRs in the ESTs revealed 1702 unique sequences containing SSRs and 1705 SSR markers were designed using those sequences. We have developed a comprehensive annotated transcriptome data set in yam to enrich the EST information in public databases. cDNA libraries were constructed from anthracnose fungus challenged leaf tissues for transcriptome characterization, and differential gene expression analysis. Thus, it helped in identifying unique transcripts in each library for disease resistance. These EST resources provide the basis for future microarray development, marker validation, genetic linkage mapping and QTL analysis in Dioscorea species.

  1. Generation and analysis of expressed sequence tags (ESTs for marker development in yam (Dioscorea alata L.

    Directory of Open Access Journals (Sweden)

    Robert Asiedu

    2011-02-01

    Full Text Available Abstract Background Anthracnose (Colletotrichum gloeosporioides is a major limiting factor in the production of yam (Dioscorea spp. worldwide. Availability of high quality sequence information is necessary for designing molecular markers associated with resistance. However, very limited sequence information pertaining to yam is available at public genome databases. Therefore, this collaborative project was developed for genetic improvement and germplasm characterization of yams using molecular markers. The current investigation is focused on studying gene expression, by large scale generation of ESTs, from one susceptible (TDa 95-0310 and two resistant yam genotypes (TDa 87-01091, TDa 95-0328 challenged with the fungus. Total RNA was isolated from young leaves of resistant and susceptible genotypes and cDNA libraries were sequenced using Roche 454 technology. Results A total of 44,757 EST sequences were generated from the cDNA libraries of the resistant and susceptible genotypes. Greater than 56% of ESTs were annotated using MapMan Mercator tool and Blast2GO search tools. Gene annotations were used to characterize the transcriptome in yam and also perform a differential gene expression analysis between the resistant and susceptible EST datasets. Mining for SSRs in the ESTs revealed 1702 unique sequences containing SSRs and 1705 SSR markers were designed using those sequences. Conclusion We have developed a comprehensive annotated transcriptome data set in yam to enrich the EST information in public databases. cDNA libraries were constructed from anthracnose fungus challenged leaf tissues for transcriptome characterization, and differential gene expression analysis. Thus, it helped in identifying unique transcripts in each library for disease resistance. These EST resources provide the basis for future microarray development, marker validation, genetic linkage mapping and QTL analysis in Dioscorea species.

  2. Carrier molecules and extraction of circulating tumor DNA for next generation sequencing in colorectal cancer.

    Science.gov (United States)

    Beránek, Martin; Sirák, Igor; Vošmik, Milan; Petera, Jiří; Drastíková, Monika; Palička, Vladimír

    The aims of the study were: i) to compare circulating tumor DNA (ctDNA) yields obtained by different manual extraction procedures, ii) to evaluate the addition of various carrier molecules into the plasma to improve ctDNA extraction recovery, and iii) to use next generation sequencing (NGS) technology to analyze KRAS, BRAF, and NRAS somatic mutations in ctDNA from patients with metastatic colorectal cancer. Venous blood was obtained from patients who suffered from metastatic colorectal carcinoma. For plasma ctDNA extraction, the following carriers were tested: carrier RNA, polyadenylic acid, glycogen, linear acrylamide, yeast tRNA, salmon sperm DNA, and herring sperm DNA. Each extract was characterized by quantitative real-time PCR and next generation sequencing. The addition of polyadenylic acid had a significant positive effect on the amount of ctDNA eluted. The sequencing data revealed five cases of ctDNA mutated in KRAS and one patient with a BRAF mutation. An agreement of 86% was found between tumor tissues and ctDNA. Testing somatic mutations in ctDNA seems to be a promising tool to monitor dynamically changing genotypes of tumor cells circulating in the body. The optimized process of ctDNA extraction should help to obtain more reliable sequencing data in patients with metastatic colorectal cancer.

  3. Characterization of the oral microbiota of healthy cats using next-generation sequencing.

    Science.gov (United States)

    Sturgeon, A; Pinder, S L; Costa, M C; Weese, J S

    2014-08-01

    The healthy feline oral cavity harbours a rich assemblage of microorganisms, which have not previously been well characterized using modern sequencing technology. The goal of this study was to accurately describe the oral microbiota of 11 healthy cats using next-generation sequencing. Sequencing generated a total of 10,177 operational taxonomic units, representing 273 genera from 18 bacterial phyla. Eight bacterial phyla made up 97.6% of sequences: Proteobacteria (75.2%), Bacteroidetes (9.3%), Firmicutes (6.7%), SR1 (2.7%), Spirochaetes (1.8%), Fusobacteria (1.3%), and Actinobacteria (0.6%). The most prevalent genus-level phylotypes were: an unclassified Pasteurellaceae (18.7%), Moraxella (10.9%), Thermomonas (6.9%), an unclassified Comamonadaceae (5.6%), Neisseria (4.9%), an unclassified Moraxellaceae (4.4%), and Pasteurella (4.3%). Results suggest that the feline oral microbiota are largely conserved between cats at the phylum level, and that the population is highly diverse, rich and even. A strong core microbiome was evident among all cats, yet significant differences in oral bacterial populations were observed across cats in each household.

  4. Implementing amplicon-based next generation sequencing in the diagnosis of small cell lung carcinoma metastases.

    Science.gov (United States)

    Meder, Lydia; König, Katharina; Fassunke, Jana; Ozretić, Luka; Wolf, Jürgen; Merkelbach-Bruse, Sabine; Heukamp, Lukas C; Buettner, Reinhard

    2015-12-01

    Small cell lung carcinoma (SCLC) is the most aggressive entity of lung cancer. Rapid cancer progression and early formation of systemic metastases drive the deadly outcome of SCLC. Recent advances in identifying oncogenes by cancer whole genome sequencing improved the understanding of SCLC carcinogenesis. However, tumor material is often limited in the clinic. Thus, it is a compulsive issue to improve SCLC diagnostics by combining established immunohistochemistry and next generation sequencing. We implemented amplicon-based next generation deep sequencing in our routine diagnostics pipeline to analyze RB1, TP53, EP300 and CREBBP, frequently mutated in SCLC. Thereby, our pipeline combined routine SCLC histology and identification of somatic mutations. We comprehensively analyzed fifty randomly collected SCLC metastases isolated from trachea and lymph nodes in comparison to specimens derived from primary SCLC. SCLC lymph node metastases showed enhanced proliferation and frequently a collapsed keratin cytoskeleton compared to SCLC metastases isolated from trachea. We identified characteristic synchronous mutations in RB1 and TP53 and non-synchronous CREBBP and EP300 mutations. Our data showed the benefit of implementing deep sequencing into routine diagnostics. We here identify oncogenic drivers and simultaneously gain further insights into SCLC tumor biology.

  5. The application of next-generation sequencing in the autozygosity mapping of human recessive diseases.

    Science.gov (United States)

    Alkuraya, Fowzan S

    2013-11-01

    Autozygosity, or the inheritance of two copies of an ancestral allele, has the potential to not only reveal phenotypes caused by biallelic mutations in autosomal recessive genes, but to also facilitate the mapping of such mutations by flagging the surrounding haplotypes as tractable runs of homozygosity (ROH), a process known as autozygosity mapping. Since SNPs replaced microsatellites as markers for the purpose of genomewide identification of ROH, autozygosity mapping of Mendelian genes has witnessed a significant acceleration. Historically, successful mapping traditionally required favorable family structure that permits the identification of an autozygous interval that is amenable to candidate gene selection and confirmation by Sanger sequencing. This requirement presented a major bottleneck that hindered the utilization of simplex cases and many multiplex families with autosomal recessive phenotypes. However, the advent of next-generation sequencing that enables massively parallel sequencing of DNA has largely bypassed this bottleneck and thus ushered in an era of unprecedented pace of Mendelian disease gene discovery. The ability to identify a single causal mutation among a massive number of variants that are uncovered by next-generation sequencing can be challenging, but applying autozygosity as a filter can greatly enhance the enrichment process and its throughput. This review will discuss the power of combining the best of both techniques in the mapping of recessive disease genes and offer some tips to troubleshoot potential limitations.

  6. Current practices and guidelines for clinical next-generation sequencing oncology testing

    Institute of Scientific and Technical Information of China (English)

    Samuel P. Strom

    2016-01-01

    Next-generation sequencing (NGS) has been rapidly integrated into molecular pathology, dramatically increasing the breadth genomic of information available to oncologists and their patients. This review will explore the ways in which this new technology is currently applied to bolster care for patients with solid tumors and hematological malignancies, focusing on practices and guidelines for assessing the technical validity and clinical utility of DNA variants identified during clinical NGS oncology testing.

  7. Characterisation of the transcriptome of a wild great tit Parus major population by next generation sequencing

    Directory of Open Access Journals (Sweden)

    Sheldon Ben C

    2011-06-01

    Full Text Available Abstract Background The recent development of next generation sequencing technologies has made it possible to generate very large amounts of sequence data in species with little or no genome information. Combined with the large phenotypic databases available for wild and non-model species, these data will provide an unprecedented opportunity to "genomicise" ecological model organisms and establish the genetic basis of quantitative traits in natural populations. Results This paper describes the sequencing, de novo assembly and analysis from the transcriptome of eight tissues of ten wild great tits. Approximately 4.6 million sequences and 1.4 billion bases of DNA were generated and assembled into 95,979 contigs, one third of which aligned with known Taeniopygia guttata (zebra finch and Gallus gallus (chicken transcripts. The majority (78% of the remaining contigs aligned within or very close to regions of the zebra finch genome containing known genes, suggesting that they represented precursor mRNA rather than untranscribed genomic DNA. More than 35,000 single nucleotide polymorphisms and 10,000 microsatellite repeats were identified. Eleven percent of contigs were expressed in every tissue, while twenty one percent of contigs were expressed in only one tissue. The function of those contigs with strong evidence for tissue specific expression and contigs expressed in every tissue was inferred from the gene ontology (GO terms associated with these contigs; heart and pancreas had the highest number of highly tissue specific GO terms (21.4% and 28.5% respectively. Conclusions In summary, the transcriptomic data generated in this study will contribute towards efforts to assemble and annotate the great tit genome, as well as providing the markers required to perform gene mapping studies in wild populations.

  8. Estimation of allele frequency and association mapping using next-generation sequencing data

    DEFF Research Database (Denmark)

    Kim, Su Yeon; Lohmueller, Kirk E; Albrechtsen, Anders;

    2011-01-01

    Estimation of allele frequency is of fundamental importance in population genetic analyses and in association mapping. In most studies using next-generation sequencing, a cost effective approach is to use medium or low-coverage data (e.g., <15X). However, SNP calling and allele frequency estimati...... in such studies is associated with substantial statistical uncertainty because of varying coverage and high error rates....

  9. Using ultra-sensitive next generation sequencing to dissect DNA damage-induced mutagenesis

    OpenAIRE

    Kaile Wang; Xiaolu Ma; Xue Zhang; Dafei Wu; Chenyi Sun; Yazhou Sun; Xuemei Lu; Chung-I Wu; Caixia Guo; Jue Ruan

    2016-01-01

    Next generation sequencing (NGS) technologies have dramatically improved studies in biology and biomedical science. However, no optimal NGS approach is available to conveniently analyze low frequency mutations caused by DNA damage treatments. Here, by developing an exquisite ultra-sensitive NGS (USNGS) platform “EasyMF” and incorporating it with a widely used supF shuttle vector-based mutagenesis system, we can conveniently dissect roles of lesion bypass polymerases in damage-induced mutagene...

  10. A second generation radiation hybrid map to aid the assembly of the bovine genome sequence

    Directory of Open Access Journals (Sweden)

    Janitz Michal

    2006-11-01

    Full Text Available Abstract Background Several approaches can be used to determine the order of loci on chromosomes and hence develop maps of the genome. However, all mapping approaches are prone to errors either arising from technical deficiencies or lack of statistical support to distinguish between alternative orders of loci. The accuracy of the genome maps could be improved, in principle, if information from different sources was combined to produce integrated maps. The publicly available bovine genomic sequence assembly with 6× coverage (Btau_2.0 is based on whole genome shotgun sequence data and limited mapping data however, it is recognised that this assembly is a draft that contains errors. Correcting the sequence assembly requires extensive additional mapping information to improve the reliability of the ordering of sequence scaffolds on chromosomes. The radiation hybrid (RH map described here has been contributed to the international sequencing project to aid this process. Results An RH map for the 30 bovine chromosomes is presented. The map was built using the Roslin 3000-rad RH panel (BovGen RH map and contains 3966 markers including 2473 new loci in addition to 262 amplified fragment-length polymorphisms (AFLP and 1231 markers previously published with the first generation RH map. Sequences of the mapped loci were aligned with published bovine genome maps to identify inconsistencies. In addition to differences in the order of loci, several cases were observed where the chromosomal assignment of loci differed between maps. All the chromosome maps were aligned with the current 6× bovine assembly (Btau_2.0 and 2898 loci were unambiguously located in the bovine sequence. The order of loci on the RH map for BTA 5, 7, 16, 22, 25 and 29 differed substantially from the assembled bovine sequence. From the 2898 loci unambiguously identified in the bovine sequence assembly, 131 mapped to different chromosomes in the BovGen RH map. Conclusion Alignment of the

  11. WIND-ACCRETION DISKS IN WIDE BINARIES, SECOND-GENERATION PROTOPLANETARY DISKS, AND ACCRETION ONTO WHITE DWARFS

    Energy Technology Data Exchange (ETDEWEB)

    Perets, Hagai B. [Technion-Israel Institute of Technology, Haifa (Israel); Kenyon, Scott J., E-mail: hperets@physics.technion.ac.il [Harvard-Smithsonian Center for Astrophysics, 60 Garden Street, Cambridge, MA 02138 (United States)

    2013-02-20

    Mass transfer from an evolved donor star to its binary companion is a standard feature of stellar evolution in binaries. In wide binaries, the companion star captures some of the mass ejected in a wind by the primary star. The captured material forms an accretion disk. Here, we study the evolution of wind-accretion disks, using a numerical approach which allows us to follow the long-term evolution. For a broad range of initial conditions, we derive the radial density and temperature profiles of the disk. In most cases, wind accretion leads to long-lived stable disks over the lifetime of the asymptotic giant branch donor star. The disks have masses of a few times 10{sup -5}-10{sup -3} M {sub Sun }, with surface density and temperature profiles that follow broken power laws. The total mass in the disk scales approximately linearly with the viscosity parameter used. Roughly, 50%-80% of the mass falling into the disk accretes onto the central star; the rest flows out through the outer edge of the disk into the stellar wind of the primary. For systems with large accretion rates, the secondary accretes as much as 0.1 M {sub Sun }. When the secondary is a white dwarf, accretion naturally leads to nova and supernova eruptions. For all types of secondary star, the surface density and temperature profiles of massive disks resemble structures observed in protoplanetary disks, suggesting that coordinated observational programs might improve our understanding of uncertain disk physics.

  12. Next generation sequencing reveals the antibiotic resistant variants in the genome of Pseudomonas aeruginosa.

    Science.gov (United States)

    Ramanathan, Babu; Jindal, Hassan Mahmood; Le, Cheng Foh; Gudimella, Ranganath; Anwar, Arif; Razali, Rozaimi; Poole-Johnson, Johan; Manikam, Rishya; Sekaran, Shamala Devi

    2017-01-01

    Rapid progress in next generation sequencing and allied computational tools have aided in identification of single nucleotide variants in genomes of several organisms. In the present study, we have investigated single nucleotide polymorphism (SNP) in ten multi-antibiotic resistant Pseudomonas aeruginosa clinical isolates. All the draft genomes were submitted to Rapid Annotations using Subsystems Technology (RAST) web server and the predicted protein sequences were used for comparison. Non-synonymous single nucleotide polymorphism (nsSNP) found in the clinical isolates compared to the reference genome (PAO1), and the comparison of nsSNPs between antibiotic resistant and susceptible clinical isolates revealed insights into the genome variation. These nsSNPs identified in the multi-drug resistant clinical isolates were found to be altering a single amino acid in several antibiotic resistant genes. We found mutations in genes encoding efflux pump systems, cell wall, DNA replication and genes involved in repair mechanism. In addition, nucleotide deletions in the genome and mutations leading to generation of stop codons were also observed in the antibiotic resistant clinical isolates. Next generation sequencing is a powerful tool to compare the whole genomes and analyse the single base pair variations found within the antibiotic resistant genes. We identified specific mutations within antibiotic resistant genes compared to the susceptible strain of the same bacterial species and these findings may provide insights to understand the role of single nucleotide variants in antibiotic resistance.

  13. Unraveling the formation history of the black hole X-ray binary LMC X-3 from the zero age main sequence to the present

    Science.gov (United States)

    Sørensen, Mads; Fragos, Tassos; Steiner, James F.; Antoniou, Vallia; Meynet, Georges; Dosopoulou, Fani

    2017-01-01

    Aims: We have endeavoured to understand the formation and evolution of the black hole (BH) X-ray binary LMC X-3. We estimated the properties of the system at four evolutionary stages: (1) at the zero-age main-sequence (ZAMS); (2) immediately before the supernova (SN) explosion of the primary; (3) immediately after the SN; and (4) at the moment when Roche-lobe overflow began. Methods: We used a hybrid approach that combined detailed calculations of the stellar structure and binary evolution with approximate population synthesis models. This allowed us to estimate potential natal kicks and the evolution of the BH spin. We incorporated as model constraints the most up-to-date observational information throughout, which include the binary orbital properties, the companion star mass, effective temperature, surface gravity and radius, and the BH mass and spin. Results: We find at 5% and 95% confidence, respectively, that LMC X-3 began as a ZAMS system with the mass of the primary star in the range M1,ZAMS = 22-31 M⊙ and a secondary star of M2,ZAMS = 5.0-8.3 M⊙, in a wide (PZAMS ≳ 2.000 days) and eccentric (eZAMS ≳ 0.18) orbit. Immediately before the SN, the primary had a mass of M1,preSN = 11.1-18.0 M⊙, but the secondary star was largely unaffected. The orbital period decreased to 0.6-1.7 days and is still eccentric 0 ≤ epreSN ≤ 0.44. We find that a symmetric SN explosion with no or small natal kicks (a few tens of km s-1) imparted on the BH cannot be formally excluded, but large natal kicks in excess of ≳120 km s-1 increase the estimated formation rate by an order of magnitude. Following the SN, the system has a BH MBH,postSN = 6.4-8.2 M⊙ and is set on an eccentric orbit. At the onset of the Roche-lobe overflow, the orbit is circular and has a period of PRLO = 0.8-1.4 days. The full Table 2 is only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (http://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/597/A12

  14. Multi-platform next-generation sequencing of the domestic turkey (Meleagris gallopavo): genome assembly and analysis

    NARCIS (Netherlands)

    Dalloul, R.A.; Long, J.A.; Zimin, A.V.; Aslam, M.L.; Crooijmans, R.P.M.A.; Megens, H.J.W.C.; Groenen, M.

    2010-01-01

    A synergistic combination of two next-generation sequencing platforms with a detailed comparative BAC physical contig map provided a cost-effective assembly of the genome sequence of the domestic turkey (Meleagris gallopavo). Heterozygosity of the sequenced source genome allowed discovery of more th

  15. Next Generation Sequencing of Actinobacteria for the Discovery of Novel Natural Products

    Directory of Open Access Journals (Sweden)

    Juan Pablo Gomez-Escribano

    2016-04-01

    Full Text Available Like many fields of the biosciences, actinomycete natural products research has been revolutionised by next-generation DNA sequencing (NGS. Hundreds of new genome sequences from actinobacteria are made public every year, many of them as a result of projects aimed at identifying new natural products and their biosynthetic pathways through genome mining. Advances in these technologies in the last five years have meant not only a reduction in the cost of whole genome sequencing, but also a substantial increase in the quality of the data, having moved from obtaining a draft genome sequence comprised of several hundred short contigs, sometimes of doubtful reliability, to the possibility of obtaining an almost complete and accurate chromosome sequence in a single contig, allowing a detailed study of gene clusters and the design of strategies for refactoring and full gene cluster synthesis. The impact that these technologies are having in the discovery and study of natural products from actinobacteria, including those from the marine environment, is only starting to be realised. In this review we provide a historical perspective of the field, analyse the strengths and limitations of the most relevant technologies, and share the insights acquired during our genome mining projects.

  16. Next Generation Sequencing of Actinobacteria for the Discovery of Novel Natural Products.

    Science.gov (United States)

    Gomez-Escribano, Juan Pablo; Alt, Silke; Bibb, Mervyn J

    2016-04-13

    Like many fields of the biosciences, actinomycete natural products research has been revolutionised by next-generation DNA sequencing (NGS). Hundreds of new genome sequences from actinobacteria are made public every year, many of them as a result of projects aimed at identifying new natural products and their biosynthetic pathways through genome mining. Advances in these technologies in the last five years have meant not only a reduction in the cost of whole genome sequencing, but also a substantial increase in the quality of the data, having moved from obtaining a draft genome sequence comprised of several hundred short contigs, sometimes of doubtful reliability, to the possibility of obtaining an almost complete and accurate chromosome sequence in a single contig, allowing a detailed study of gene clusters and the design of strategies for refactoring and full gene cluster synthesis. The impact that these technologies are having in the discovery and study of natural products from actinobacteria, including those from the marine environment, is only starting to be realised. In this review we provide a historical perspective of the field, analyse the strengths and limitations of the most relevant technologies, and share the insights acquired during our genome mining projects.

  17. Performance Characteristics and Validation of Next-Generation Sequencing for Human Leucocyte Antigen Typing.

    Science.gov (United States)

    Weimer, Eric T; Montgomery, Maureen; Petraroia, Rosanne; Crawford, John; Schmitz, John L

    2016-09-01

    High-resolution human leukocyte antigen (HLA) matching reduces graft-versus-host disease and improves overall patient survival after hematopoietic stem cell transplant. Sanger sequencing has been the gold standard for HLA typing since 1996. However, given the increasing number of new HLA alleles identified and the complexity of the HLA genes, clinical HLA typing by Sanger sequencing requires several rounds of additional testing to provide allele-level resolution. Although next-generation sequencing (NGS) is routinely used in molecular genetics, few clinical HLA laboratories use the technology. The performance characteristics of NGS HLA typing using TruSight HLA were determined using Sanger sequencing as the reference method. In total, 211 samples were analyzed with an overall accuracy of 99.8% (2954/2961) and 46 samples were analyzed for precision with 100% (368/368) reproducibility. Most discordant alleles were because of technical error rather than assay performance. More important, the ambiguity rate was 3.5% (103/2961). Seventy-four percentage of the ambiguities were within the DRB1 and DRB4 loci. HLA typing by NGS saves approximately $6000 per run when compared to Sanger sequencing. Thus, TruSight HLA assay enables high-throughput HLA typing with an accuracy, precision, ambiguity rate, and cost savings that should facilitate adoption of NGS technology in clinical HLA laboratories.

  18. Next-Generation Sequencing in Clinical Molecular Diagnostics of Cancer: Advantages and Challenges

    Directory of Open Access Journals (Sweden)

    Rajyalakshmi Luthra

    2015-10-01

    Full Text Available The application of next-generation sequencing (NGS to characterize cancer genomes has resulted in the discovery of numerous genetic markers. Consequently, the number of markers that warrant routine screening in molecular diagnostic laboratories, often from limited tumor material, has increased. This increased demand has been difficult to manage by traditional low- and/or medium-throughput sequencing platforms. Massively parallel sequencing capabilities of NGS provide a much-needed alternative for mutation screening in multiple genes with a single low investment of DNA. However, implementation of NGS technologies, most of which are for research use only (RUO, in a diagnostic laboratory, needs extensive validation in order to establish Clinical Laboratory Improvement Amendments (CLIA and College of American Pathologists (CAP-compliant performance characteristics. Here, we have reviewed approaches for validation of NGS technology for routine screening of tumors. We discuss the criteria for selecting gene markers to include in the NGS panel and the deciding factors for selecting target capture approaches and sequencing platforms. We also discuss challenges in result reporting, storage and retrieval of the voluminous sequencing data and the future potential of clinical NGS.

  19. Novel Primer Sets for Next Generation Sequencing-Based Analyses of Water Quality.

    Science.gov (United States)

    Lee, Elvina; Khurana, Maninder S; Whiteley, Andrew S; Monis, Paul T; Bath, Andrew; Gordon, Cameron; Ryan, Una M; Paparini, Andrea

    2017-01-01

    Next generation sequencing (NGS) has rapidly become an invaluable tool for the detection, identification and relative quantification of environmental microorganisms. Here, we demonstrate two new 16S rDNA primer sets, which are compatible with NGS approaches and are primarily for use in water quality studies. Compared to 16S rRNA gene based universal primers, in silico and experimental analyses demonstrated that the new primers showed increased specificity for the Cyanobacteria and Proteobacteria phyla, allowing increased sensitivity for the detection, identification and relative quantification of toxic bloom-forming microalgae, microbial water quality bioindicators and common pathogens. Significantly, Cyanobacterial and Proteobacterial sequences accounted for ca. 95% of all sequences obtained within NGS runs (when compared to ca. 50% with standard universal NGS primers), providing higher sensitivity and greater phylogenetic resolution of key water quality microbial groups. The increased selectivity of the new primers allow the parallel sequencing of more samples through reduced sequence retrieval levels required to detect target groups, potentially reducing NGS costs by 50% but still guaranteeing optimal coverage and species discrimination.

  20. Next-generation sequencing analysis for detecting human papillomavirus in oral verrucous carcinoma.

    Science.gov (United States)

    Samman, Manar; Wood, Henry; Conway, Caroline; Berri, Stefano; Pentenero, Monica; Gandolfo, Sergio; Cassenti, Adele; Cassoni, Paola; Al Ajlan, Abdulaziz; Barrett, A William; Chengot, Preetha; MacLennan, Kenneth; High, Alec S; Rabbitts, Pamela

    2014-07-01

    The etiology of oral verrucous carcinoma is unknown, and human papillomavirus 'involvement' remains contentious. The uncertainty can be attributed to varied detection procedures and difficulties in defining 'gold-standard' histologic criteria for diagnosing 'verrucous' lesions. Their paucity also hampers investigation. We aimed to analyze oral verrucous lesions for human papillomavirus (HPV) subtype genomes. We used next-generation sequencing for the detection of papillomavirus sequences, identifying subtypes and computing viral loads. We identified a total of 78 oral verrucous cases (62 carcinomas and 16 hyperplasias). DNA was extracted from all and sequenced at a coverage between 2.5% and 13%. An HPV-16 sequence was detected in 1 carcinoma and 1 hyperplasia, and an HPV-2 sequence was detected in 1 carcinoma out of the 78 cases, with viral loads of 2.24, 8.16, and 0.33 viral genomes per cell, respectively. Our results indicate no conclusive human papillomavirus involvement in oral verrucous carcinoma or hyperplasia. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Analysis of plant microbe interactions in the era of next generation sequencing technologies

    Directory of Open Access Journals (Sweden)

    Claudia eKnief

    2014-05-01

    Full Text Available Next generation sequencing (NGS technologies have impressively accelerated research in biological science during the last years by enabling the production of large volumes of sequence data to a drastically lower price per base, compared to traditional sequencing methods. The recent and ongoing developments in the field allow addressing research questions in plant-microbe biology that were not conceivable just a few years ago. The present review provides an overview of NGS technologies and their usefulness for the analysis of microorganisms that live in association with plants. Possible limitations of the different sequencing systems, in particular sources of errors and bias, are critically discussed and methods are disclosed that help to overcome these shortcomings. A focus will be on the application of NGS methods in metagenomic studies, including the analysis of microbial communities by amplicon sequencing, which can be considered as a targeted metagenomic approach. Different applications of NGS technologies are exemplified by selected research articles that address the biology of the pant associated microbiota to demonstrate the worth of the new methods.

  2. Paging through history: parchment as a reservoir of ancient DNA for next generation sequencing.

    Science.gov (United States)

    Teasdale, M D; van Doorn, N L; Fiddyment, S; Webb, C C; O'Connor, T; Hofreiter, M; Collins, M J; Bradley, D G

    2015-01-19

    Parchment represents an invaluable cultural reservoir. Retrieving an additional layer of information from these abundant, dated livestock-skins via the use of ancient DNA (aDNA) sequencing has been mooted by a number of researchers. However, prior PCR-based work has indicated that this may be challenged by cross-individual and cross-species contamination, perhaps from the bulk parchment preparation process. Here we apply next generation sequencing to two parchments of seventeenth and eighteenth century northern English provenance. Following alignment to the published sheep, goat, cow and human genomes, it is clear that the only genome displaying substantial unique homology is sheep and this species identification is confirmed by collagen peptide mass spectrometry. Only 4% of sequence reads align preferentially to a different species indicating low contamination across species. Moreover, mitochondrial DNA sequences suggest an upper bound of contamination at 5%. Over 45% of reads aligned to the sheep genome, and even this limited sequencing exercise yield 9 and 7% of each sampled sheep genome post filtering, allowing the mapping of genetic affinity to modern British sheep breeds. We conclude that parchment represents an excellent substrate for genomic analyses of historical livestock.

  3. ParticleCall: A particle filter for base calling in next-generation sequencing systems

    Directory of Open Access Journals (Sweden)

    Shen Xiaohu

    2012-07-01

    Full Text Available Abstract Background Next-generation sequencing systems are capable of rapid and cost-effective DNA sequencing, thus enabling routine sequencing tasks and taking us one step closer to personalized medicine. Accuracy and lengths of their reads, however, are yet to surpass those provided by the conventional Sanger sequencing method. This motivates the search for computationally efficient algorithms capable of reliable and accurate detection of the order of nucleotides in short DNA fragments from the acquired data. Results In this paper, we consider Illumina’s sequencing-by-synthesis platform which relies on reversible terminator chemistry and describe the acquired signal by reformulating its mathematical model as a Hidden Markov Model. Relying on this model and sequential Monte Carlo methods, we develop a parameter estimation and base calling scheme called ParticleCall. ParticleCall is tested on a data set obtained by sequencing phiX174 bacteriophage using Illumina’s Genome Analyzer II. The results show that the developed base calling scheme is significantly more computationally efficient than the best performing unsupervised method currently available, while achieving the same accuracy. Conclusions The proposed ParticleCall provides more accurate calls than the Illumina’s base calling algorithm, Bustard. At the same time, ParticleCall is significantly more computationally efficient than other recent schemes with similar performance, rendering it more feasible for high-throughput sequencing data analysis. Improvement of base calling accuracy will have immediate beneficial effects on the performance of downstream applications such as SNP and genotype calling. ParticleCall is freely available at https://sourceforge.net/projects/particlecall.

  4. BG7: A New Approach for Bacterial Genome Annotation Designed for Next Generation Sequencing Data

    Science.gov (United States)

    Pareja-Tobes, Pablo; Manrique, Marina; Pareja-Tobes, Eduardo; Pareja, Eduardo; Tobes, Raquel

    2012-01-01

    BG7 is a new system for de novo bacterial, archaeal and viral genome annotation based on a new approach specifically designed for annotating genomes sequenced with next generation sequencing technologies. The system is versatile and able to annotate genes even in the step of preliminary assembly of the genome. It is especially efficient detecting unexpected genes horizontally acquired from bacterial or archaeal distant genomes, phages, plasmids, and mobile elements. From the initial phases of the gene annotation process, BG7 exploits the massive availability of annotated protein sequences in databases. BG7 predicts ORFs and infers their function based on protein similarity with a wide set of reference proteins, integrating ORF prediction and functional annotation phases in just one step. BG7 is especially tolerant to sequencing errors in start and stop codons, to frameshifts, and to assembly or scaffolding errors. The system is also tolerant to the high level of gene fragmentation which is frequently found in not fully assembled genomes. BG7 current version – which is developed in Java, takes advantage of Amazon Web Services (AWS) cloud computing features, but it can also be run locally in any operating system. BG7 is a fast, automated and scalable system that can cope with the challenge of analyzing the huge amount of genomes that are being sequenced with NGS technologies. Its capabilities and efficiency were demonstrated in the 2011 EHEC Germany outbreak in which BG7 was used to get the first annotations right the next day after the first entero-hemorrhagic E. coli genome sequences were made publicly available. The suitability of BG7 for genome annotation has been proved for Illumina, 454, Ion Torrent, and PacBio sequencing technologies. Besides, thanks to its plasticity, our system could be very easily adapted to work with new technologies in the future. PMID:23185310

  5. Mutation Scanning in Wheat by Exon Capture and Next-Generation Sequencing.

    Directory of Open Access Journals (Sweden)

    Robert King

    Full Text Available Targeted Induced Local Lesions in Genomes (TILLING is a reverse genetics approach to identify novel sequence variation in genomes, with the aims of investigating gene function and/or developing useful alleles for breeding. Despite recent advances in wheat genomics, most current TILLING methods are low to medium in throughput, being based on PCR amplification of the target genes. We performed a pilot-scale evaluation of TILLING in wheat by next-generation sequencing through exon capture. An oligonucleotide-based enrichment array covering ~2 Mbp of wheat coding sequence was used to carry out exon capture and sequencing on three mutagenised lines of wheat containing previously-identified mutations in the TaGA20ox1 homoeologous genes. After testing different mapping algorithms and settings, candidate SNPs were identified by mapping to the IWGSC wheat Chromosome Survey Sequences. Where sequence data for all three homoeologues were found in the reference, mutant calls were unambiguous; however, where the reference lacked one or two of the homoeologues, captured reads from these genes were mis-mapped to other homoeologues, resulting either in dilution of the variant allele frequency or assignment of mutations to the wrong homoeologue. Competitive PCR assays were used to validate the putative SNPs and estimate cut-off levels for SNP filtering. At least 464 high-confidence SNPs were detected across the three mutagenized lines, including the three known alleles in TaGA20ox1, indicating a mutation rate of ~35 SNPs per Mb, similar to that estimated by PCR-based TILLING. This demonstrates the feasibility of using exon capture for genome re-sequencing as a method of mutation detection in polyploid wheat, but accurate mutation calling will require an improved genomic reference with more comprehensive coverage of homoeologues.

  6. Combinatorial analysis and algorithms for quasispecies reconstruction using next-generation sequencing

    Directory of Open Access Journals (Sweden)

    Vincenti Donatella

    2011-01-01

    Full Text Available Abstract Background Next-generation sequencing (NGS offers a unique opportunity for high-throughput genomics and has potential to replace Sanger sequencing in many fields, including de-novo sequencing, re-sequencing, meta-genomics, and characterisation of infectious pathogens, such as viral quasispecies. Although methodologies and software for whole genome assembly and genome variation analysis have been developed and refined for NGS data, reconstructing a viral quasispecies using NGS data remains a challenge. This application would be useful for analysing intra-host evolutionary pathways in relation to immune responses and antiretroviral therapy exposures. Here we introduce a set of formulae for the combinatorial analysis of a quasispecies, given a NGS re-sequencing experiment and an algorithm for quasispecies reconstruction. We require that sequenced fragments are aligned against a reference genome, and that the reference genome is partitioned into a set of sliding windows (amplicons. The reconstruction algorithm is based on combinations of multinomial distributions and is designed to minimise the reconstruction of false variants, called in-silico recombinants. Results The reconstruction algorithm was applied to error-free simulated data and reconstructed a high percentage of true variants, even at a low genetic diversity, where the chance to obtain in-silico recombinants is high. Results on empirical NGS data from patients infected with hepatitis B virus, confirmed its ability to characterise different viral variants from distinct patients. Conclusions The combinatorial analysis provided a description of the difficulty to reconstruct a quasispecies, given a determined amplicon partition and a measure of population diversity. The reconstruction algorithm showed good performance both considering simulated data and real data, even in presence of sequencing errors.

  7. Exome sequencing generates high quality data in non-target regions

    Directory of Open Access Journals (Sweden)

    Guo Yan

    2012-05-01

    Full Text Available Abstract Background Exome sequencing using next-generation sequencing technologies is a cost efficient approach to selectively sequencing coding regions of human genome for detection of disease variants. A significant amount of DNA fragments from the capture process fall outside target regions, and sequence data for positions outside target regions have been mostly ignored after alignment. Result We performed whole exome sequencing on 22 subjects using Agilent SureSelect capture reagent and 6 subjects using Illumina TrueSeq capture reagent. We also downloaded sequencing data for 6 subjects from the 1000 Genomes Project Pilot 3 study. Using these data, we examined the quality of SNPs detected outside target regions by computing consistency rate with genotypes obtained from SNP chips or the Hapmap database, transition-transversion (Ti/Tv ratio, and percentage of SNPs inside dbSNP. For all three platforms, we obtained high-quality SNPs outside target regions, and some far from target regions. In our Agilent SureSelect data, we obtained 84,049 high-quality SNPs outside target regions compared to 65,231 SNPs inside target regions (a 129% increase. For our Illumina TrueSeq data, we obtained 222,171 high-quality SNPs outside target regions compared to 95,818 SNPs inside target regions (a 232% increase. For the data from the 1000 Genomes Project, we obtained 7,139 high-quality SNPs outside target regions compared to 1,548 SNPs inside target regions (a 461% increase. Conclusions These results demonstrate that a significant amount of high quality genotypes outside target regions can be obtained from exome sequencing data. These data should not be ignored in genetic epidemiology studies.

  8. 454 next generation-sequencing outperforms allele-specific PCR, Sanger sequencing, and pyrosequencing for routine KRAS mutation analysis of formalin-fixed, paraffin-embedded samples.

    Science.gov (United States)

    Altimari, Annalisa; de Biase, Dario; De Maglio, Giovanna; Gruppioni, Elisa; Capizzi, Elisa; Degiovanni, Alessio; D'Errico, Antonia; Pession, Annalisa; Pizzolitto, Stefano; Fiorentino, Michelangelo; Tallini, Giovanni

    2013-01-01

    Detection of KRAS mutations in archival pathology samples is critical for therapeutic appropriateness of anti-EGFR monoclonal antibodies in colorectal cancer. We compared the sensitivity, specificity, and accuracy of Sanger sequencing, ARMS-Scorpion (TheraScreen®) real-time polymerase chain reaction (PCR), pyrosequencing, chip array hybridization, and 454 next-generation sequencing to assess KRAS codon 12 and 13 mutations in 60 nonconsecutive selected cases of colorectal cancer. Twenty of the 60 cases were detected as wild-type KRAS by all methods with 100% specificity. Among the 40 mutated cases, 13 were discrepant with at least one method. The sensitivity was 85%, 90%, 93%, and 92%, and the accuracy was 90%, 93%, 95%, and 95% for Sanger sequencing, TheraScreen real-time PCR, pyrosequencing, and chip array hybridization, respectively. The main limitation of Sanger sequencing was its low analytical sensitivity, whereas TheraScreen real-time PCR, pyrosequencing, and chip array hybridization showed higher sensitivity but suffered from the limitations of predesigned assays. Concordance between the methods was k = 0.79 for Sanger sequencing and k > 0.85 for the other techniques. Tumor cell enrichment correlated significantly with the abundance of KRAS-mutated deoxyribonucleic acid (DNA), evaluated as ΔCt for TheraScreen real-time PCR (P = 0.03), percentage of mutation for pyrosequencing (P = 0.001), ratio for chip array hybridization (P = 0.003), and percentage of mutation for 454 next-generation sequencing (P = 0.004). Also, 454 next-generation sequencing showed the best cross correlation for quantification of mutation abundance compared with all the other methods (P < 0.001). Our comparison showed the superiority of next-generation sequencing over the other techniques in terms of sensitivity and specificity. Next-generation sequencing will replace Sanger sequencing as the reference technique for diagnostic detection of KRAS mutation in archival tumor tissues.

  9. Analysis of Litopenaeus vannamei transcriptome using the next-generation DNA sequencing technique.

    Directory of Open Access Journals (Sweden)

    Chaozheng Li

    Full Text Available BACKGROUND: Pacific white shrimp (Litopenaeus vannamei, the major species of farmed shrimps in the world, has been attracting extensive studies, which require more and more genome background knowledge. The now available transcriptome data of L. vannamei are insufficient for research requirements, and have not been adequately assembled and annotated. METHODOLOGY/PRINCIPAL FINDINGS: This is the first study that used a next-generation high-throughput DNA sequencing technique, the Solexa/Illumina GA II method, to analyze the transcriptome from whole bodies of L. vannamei larvae. More than 2.4 Gb of raw data were generated, and 109,169 unigenes with a mean length of 396 bp were assembled using the SOAP denovo software. 73,505 unigenes (>200 bp with good quality sequences were selected and subjected to annotation analysis, among which 37.80% can be matched in NCBI Nr database, 37.3% matched in Swissprot, and 44.1% matched in TrEMBL. Using BLAST and BLAST2Go softwares, 11,153 unigenes were classified into 25 Clusters of Orthologous Groups of proteins (COG categories, 8171 unigenes were assigned into 51 Gene ontology (GO functional groups, and 18,154 unigenes were divided into 220 Kyoto Encyclopedia of Genes and Genomes (KEGG pathways. To primarily verify part of the results of assembly and annotations, 12 assembled unigenes that are homologous to many embryo development-related genes were chosen and subjected to RT-PCR for electrophoresis and Sanger sequencing analyses, and to real-time PCR for expression profile analyses during embryo development. CONCLUSIONS/SIGNIFICANCE: The L. vannamei transcriptome analyzed using the next-generation sequencing technique enriches the information of L. vannamei genes, which will facilitate our understanding of the genome background of crustaceans, and promote the studies on L. vannamei.

  10. High-resolution analysis of the 5'-end transcriptome using a next generation DNA sequencer.

    Directory of Open Access Journals (Sweden)

    Shin-ichi Hashimoto

    Full Text Available Massively parallel, tag-based sequencing systems, such as the SOLiD system, hold the promise of revolutionizing the study of whole genome gene expression due to the number of data points that can be generated in a simple and cost-effective manner. We describe the development of a 5'-end transcriptome workflow for the SOLiD system and demonstrate the advantages in sensitivity and dynamic range offered by this tag-based application over traditional approaches for the study of whole genome gene expression. 5'-end transcriptome analysis was used to study whole genome gene expression within a colon cancer cell line, HT-29, treated with the DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5Aza. More than 20 million 25-base 5'-end tags were obtained from untreated and 5Aza-treated cells and matched to sequences within the human genome. Seventy three percent of the mapped unique tags were associated with RefSeq cDNA sequences, corresponding to approximately 14,000 different protein-coding genes in this single cell type. The level of expression of these genes ranged from 0.02 to 4,704 transcripts per cell. The sensitivity of a single sequence run of the SOLiD platform was 100-1,000 fold greater than that observed from 5'end SAGE data generated from the analysis of 70,000 tags obtained by Sanger sequencing. The high-resolution 5'end gene expression profiling presented in this study will not only provide novel insight into the transcriptional machinery but should also serve as a basis for a better understanding of cell biology.

  11. Next-generation sequencing traces human induced pluripotent stem cell lines clonally generated from heterogeneous cancer tissue.

    Science.gov (United States)

    Ishikawa, Tetsuya

    2017-05-26

    To investigate genotype variation among induced pluripotent stem cell (iPSC) lines that were clonally generated from heterogeneous colon cancer tissues using next-generation sequencing. Human iPSC lines were clonally established by selecting independent single colonies expanded from heterogeneous primary cells of S-shaped colon cancer tissues by retroviral gene transfer (OCT3/4, SOX2, and KLF4). The ten iPSC lines, their starting cancer tissues, and the matched adjacent non-cancerous tissues were analyzed using next-generation sequencing and bioinformatics analysis using the human reference genome hg19. Non-synonymous single-nucleotide variants (SNVs) (missense, nonsense, and read-through) were identified within the target region of 612 genes related to cancer and the human kinome. All SNVs were annotated using dbSNP135, CCDS, RefSeq, GENCODE, and 1000 Genomes. The SNVs of the iPSC lines were compared with the genotypes of the cancerous and non-cancerous tissues. The putative genotypes were validated using allelic depth and genotype quality. For final confirmation, mutated genotypes were manually curated using the Integrative Genomics Viewer. In eight of the ten iPSC lines, one or two non-synonymous SNVs in EIF2AK2, TTN, ULK4, TSSK1B, FLT4, STK19, STK31, TRRAP, WNK1, PLK1 or PIK3R5 were identified as novel SNVs and were not identical to the genotypes found in the cancer and non-cancerous tissues. This result suggests that the SNVs were de novo or pre-existing mutations that originated from minor populations, such as multifocal pre-cancer (stem) cells or pre-metastatic cancer cells from multiple, different clonal evolutions, present within the heterogeneous cancer tissue. The genotypes of all ten iPSC lines were different from the mutated ERBB2 and MKNK2 genotypes of the cancer tissues and were identical to those of the non-cancerous tissues and that found in the human reference genome hg19. Furthermore, two of the ten iPSC lines did not have any confirmed mutated

  12. Calculation of Tajima's D and other neutrality test statistics from low depth next-generation sequencing data

    DEFF Research Database (Denmark)

    Korneliussen, Thorfinn Sand; Moltke, Ida; Albrechtsen, Anders

    2013-01-01

    A number of different statistics are used for detecting natural selection using DNA sequencing data, including statistics that are summaries of the frequency spectrum, such as Tajima's D. These statistics are now often being applied in the analysis of Next Generation Sequencing (NGS) data. However......, estimates of frequency spectra from NGS data are strongly affected by low sequencing coverage; the inherent technology dependent variation in sequencing depth causes systematic differences in the value of the statistic among genomic regions....

  13. Efficient generation of cavitation bubbles and reactive oxygen species using triggered high-intensity focused ultrasound sequence for sonodynamic treatment

    Science.gov (United States)

    Yasuda, Jun; Yoshizawa, Shin; Umemura, Shin-ichiro

    2016-07-01

    Sonodynamic treatment is a method of treating cancer using reactive oxygen species (ROS) generated by cavitation bubbles in collaboration with a sonosensitizer at a target tissue. In this treatment method, both localized ROS generation and ROS generation with high efficiency are important. In this study, a triggered high-intensity focused ultrasound (HIFU) sequence, which consists of a short, extremely high intensity pulse immediately followed by a long, moderate-intensity burst, was employed for the efficient generation of ROS. In experiments, a solution sealed in a chamber was exposed to a triggered HIFU sequence. Then, the distribution of generated ROS was observed by the luminol reaction, and the amount of generated ROS was quantified using KI method. As a result, the localized ROS generation was demonstrated by light emission from the luminol reaction. Moreover, it was demonstrated that the triggered HIFU sequence has higher efficiency of ROS generation by both the KI method and the luminol reaction emission.

  14. Next Generation Sequence Analysis and Computational Genomics Using Graphical Pipeline Workflows

    Directory of Open Access Journals (Sweden)

    Marquis P. Vawter

    2012-08-01

    Full Text Available Whole-genome and exome sequencing have already proven to be essential and powerful methods to identify genes responsible for simple Mendelian inherited disorders. These methods can be applied to complex disorders as well, and have been adopted as one of the current mainstream approaches in population genetics. These achievements have been made possible by next generation sequencing (NGS technologies, which require substantial bioinformatics resources to analyze the dense and complex sequence data. The huge analytical burden of data from genome sequencing might be seen as a bottleneck slowing the publication of NGS papers at this time, especially in psychiatric genetics. We review the existing methods for processing NGS data, to place into context the rationale for the design of a computational resource. We describe our method, the Graphical Pipeline for Computational Genomics (GPCG, to perform the computational steps required to analyze NGS data. The GPCG implements flexible workflows for basic sequence alignment, sequence data quality control, single nucleotide polymorphism analysis, copy number variant identification, annotation, and visualization of results. These workflows cover all the analytical steps required for NGS data, from processing the raw reads to variant calling and annotation. The current version of the pipeline is freely available at http://pipeline.loni.ucla.edu. These applications of NGS analysis may gain clinical utility in the near future (e.g., identifying miRNA signatures in diseases when the bioinformatics approach is made feasible. Taken together, the annotation tools and strategies that have been developed to retrieve information and test hypotheses about the functional role of variants present in the human genome will help to pinpoint the genetic risk factors for psychiatric disorders.

  15. Use of four next-generation sequencing platforms to determine HIV-1 coreceptor tropism.

    Science.gov (United States)

    Archer, John; Weber, Jan; Henry, Kenneth; Winner, Dane; Gibson, Richard; Lee, Lawrence; Paxinos, Ellen; Arts, Eric J; Robertson, David L; Mimms, Larry; Quiñones-Mateu, Miguel E

    2012-01-01

    HIV-1 coreceptor tropism assays are required to rule out the presence of CXCR4-tropic (non-R5) viruses prior treatment with CCR5 antagonists. Phenotypic (e.g., Trofile™, Monogram Biosciences) and genotypic (e.g., population sequencing linked to bioinformatic algorithms) assays are the most widely used. Although several next-generation sequencing (NGS) platforms are available, to date all published deep sequencing HIV-1 tropism studies have used the 454™ Life Sciences/Roche platform. In this study, HIV-1 co-receptor usage was predicted for twelve patients scheduled to start a maraviroc-based antiretroviral regimen. The V3 region of the HIV-1 env gene was sequenced using four NGS platforms: 454™, PacBio® RS (Pacific Biosciences), Illumina®, and Ion Torrent™ (Life Technologies). Cross-platform variation was evaluated, including number of reads, read length and error rates. HIV-1 tropism was inferred using Geno2Pheno, Web PSSM, and the 11/24/25 rule and compared with Trofile™ and virologic response to antiretroviral therapy. Error rates related to insertions/deletions (indels) and nucleotide substitutions introduced by the four NGS platforms were low compared to the actual HIV-1 sequence variation. Each platform detected all major virus variants within the HIV-1 population with similar frequencies. Identification of non-R5 viruses was comparable among the four platforms, with minor differences attributable to the algorithms used to infer HIV-1 tropism. All NGS platforms showed similar concordance with virologic response to the maraviroc-based regimen (75% to 80% range depending on the algorithm used), compared to Trofile (80%) and population sequencing (70%). In conclusion, all four NGS platforms were able to detect minority non-R5 variants at comparable levels suggesting that any NGS-based method can be used to predict HIV-1 coreceptor usage.

  16. Next generation sequence analysis and computational genomics using graphical pipeline workflows.

    Science.gov (United States)

    Torri, Federica; Dinov, Ivo D; Zamanyan, Alen; Hobel, Sam; Genco, Alex; Petrosyan, Petros; Clark, Andrew P; Liu, Zhizhong; Eggert, Paul; Pierce, Jonathan; Knowles, James A; Ames, Joseph; Kesselman, Carl; Toga, Arthur W; Potkin, Steven G; Vawter, Marquis P; Macciardi, Fabio

    2012-08-30

    Whole-genome and exome sequencing have already proven to be essential and powerful methods to identify genes responsible for simple Mendelian inherited disorders. These methods can be applied to complex disorders as well, and have been adopted as one of the current mainstream approaches in population genetics. These achievements have been made possible by next generation sequencing (NGS) technologies, which require substantial bioinformatics resources to analyze the dense and complex sequence data. The huge analytical burden of data from genome sequencing might be seen as a bottleneck slowing the publication of NGS papers at this time, especially in psychiatric genetics. We review the existing methods for processing NGS data, to place into context the rationale for the design of a computational resource. We describe our method, the Graphical Pipeline for Computational Genomics (GPCG), to perform the computational steps required to analyze NGS data. The GPCG implements flexible workflows for basic sequence alignment, sequence data quality control, single nucleotide polymorphism analysis, copy number variant identification, annotation, and visualization of results. These workflows cover all the analytical steps required for NGS data, from processing the raw reads to variant calling and annotation. The current version of the pipeline is freely available at http://pipeline.loni.ucla.edu. These applications of NGS analysis may gain clinical utility in the near future (e.g., identifying miRNA signatures in diseases) when the bioinformatics approach is made feasible. Taken together, the annotation tools and strategies that have been developed to retrieve information and test hypotheses about the functional role of variants present in the human genome will help to pinpoint the genetic risk factors for psychiatric disorders.

  17. Next-generation sequencing for rodent barcoding: species identification from fresh, degraded and environmental samples.

    Science.gov (United States)

    Galan, Maxime; Pagès, Marie; Cosson, Jean-François

    2012-01-01

    Rodentia is the most diverse order among mammals, with more than 2,000 species currently described. Most of the time, species assignation is so difficult based on morphological data solely that identifying rodents at the specific level corresponds to a real challenge. In this study, we compared the applicability of 100 bp mini-barcodes from cytochrome b and cytochrome c oxidase 1 genes to enable rodent species identification. Based on GenBank sequence datasets of 115 rodent species, a 136 bp fragment of cytochrome b was selected as the most discriminatory mini-barcode, and rodent universal primers surrounding this fragment were designed. The efficacy of this new molecular tool was assessed on 946 samples including rodent tissues, feces, museum samples and feces/pellets from predators known to ingest rodents. Utilizing next-generation sequencing technologies able to sequence mixes of DNA, 1,140 amplicons were tagged, multiplexed and sequenced together in one single 454 GS-FLX run. Our method was initially validated on a reference sample set including 265 clearly identified rodent tissues, corresponding to 103 different species. Following validation, 85.6% of 555 rodent samples from Europe, Asia and Africa whose species identity was unknown were able to be identified using the BLASTN program and GenBank reference sequences. In addition, our method proved effective even on degraded rodent DNA samples: 91.8% and 75.9% of samples from feces and museum specimens respectively were correctly identified. Finally, we succeeded in determining the diet of 66.7% of the investigated carnivores from their feces and 81.8% of owls from their pellets. Non-rodent species were also identified, suggesting that our method is sensitive enough to investigate complete predator diets. This study demonstrates how this molecular identification method combined with high-throughput sequencing can open new realms of possibilities in achieving fast, accurate and inexpensive species identification.

  18. Use of four next-generation sequencing platforms to determine HIV-1 coreceptor tropism.

    Directory of Open Access Journals (Sweden)

    John Archer

    Full Text Available HIV-1 coreceptor tropism assays are required to rule out the presence of CXCR4-tropic (non-R5 viruses prior treatment with CCR5 antagonists. Phenotypic (e.g., Trofile™, Monogram Biosciences and genotypic (e.g., population sequencing linked to bioinformatic algorithms assays are the most widely used. Although several next-generation sequencing (NGS platforms are available, to date all published deep sequencing HIV-1 tropism studies have used the 454™ Life Sciences/Roche platform. In this study, HIV-1 co-receptor usage was predicted for twelve patients scheduled to start a maraviroc-based antiretroviral regimen. The V3 region of the HIV-1 env gene was sequenced using four NGS platforms: 454™, PacBio® RS (Pacific Biosciences, Illumina®, and Ion Torrent™ (Life Technologies. Cross-platform variation was evaluated, including number of reads, read length and error rates. HIV-1 tropism was inferred using Geno2Pheno, Web PSSM, and the 11/24/25 rule and compared with Trofile™ and virologic response to antiretroviral therapy. Error rates related to insertions/deletions (indels and nucleotide substitutions introduced by the four NGS platforms were low compared to the actual HIV-1 sequence variation. Each platform detected all major virus variants within the HIV-1 population with similar frequencies. Identification of non-R5 viruses was comparable among the four platforms, with minor differences attributable to the algorithms used to infer HIV-1 tropism. All NGS platforms showed similar concordance with virologic response to the maraviroc-based regimen (75% to 80% range depending on the algorithm used, compared to Trofile (80% and population sequencing (70%. In conclusion, all four NGS platforms were able to detect minority non-R5 variants at comparable levels suggesting that any NGS-based method can be used to predict HIV-1 coreceptor usage.

  19. ngs_backbone: a pipeline for read cleaning, mapping and SNP calling using Next Generation Sequence

    Directory of Open Access Journals (Sweden)

    Cañizares Joaquin

    2011-06-01

    Full Text Available Abstract Background The possibilities offered by next generation sequencing (NGS platforms are revolutionizing biotechnological laboratories. Moreover, the combination of NGS sequencing and affordable high-throughput genotyping technologies is facilitating the rapid discovery and use of SNPs in non-model species. However, this abundance of sequences and polymorphisms creates new software needs. To fulfill these needs, we have developed a powerful, yet easy-to-use application. Results The ngs_backbone software is a parallel pipeline capable of analyzing Sanger, 454, Illumina and SOLiD (Sequencing by Oligonucleotide Ligation and Detection sequence reads. Its main supported analyses are: read cleaning, transcriptome assembly and annotation, read mapping and single nucleotide polymorphism (SNP calling and selection. In order to build a truly useful tool, the software development was paired with a laboratory experiment. All public tomato Sanger EST reads plus 14.2 million Illumina reads were employed to test the tool and predict polymorphism in tomato. The cleaned reads were mapped to the SGN tomato transcriptome obtaining a coverage of 4.2 for Sanger and 8.5 for Illumina. 23,360 single nucleotide variations (SNVs were predicted. A total of 76 SNVs were experimentally validated, and 85% were found to be real. Conclusions ngs_backbone is a new software package capable of analyzing sequences produced by NGS technologies and predicting SNVs with great accuracy. In our tomato example, we created a highly polymorphic collection of SNVs that will be a useful resource for tomato researchers and breeders. The software developed along with its documentation is freely available under the AGPL license and can be downloaded from http://bioinf.comav.upv.es/ngs_backbone/ or http://github.com/JoseBlanca/franklin.

  20. Next generation sequencing and omics in cucumber (Cucumis sativus L.) breeding directed research.

    Science.gov (United States)

    Pawełkowicz, Magdalena; Zieliński, Konrad; Zielińska, Dorota; Pląder, Wojciech; Yagi, Kouhei; Wojcieszek, Michał; Siedlecka, Ewa; Bartoszewski, Grzegorz; Skarzyńska, Agnieszka; Przybecki, Zbigniew

    2016-01-01

    In the post-genomic era the availability of genomic tools and resources is leading us to novel generation methods in plant breeding, as they facilitate the study of the genotype and its relationship with the phenotype, in particular for complex traits. In this study we have mainly concentrated on the Cucumis sativus and (but much less) Cucurbitaceae family several important vegetable crops. There are many reports on research conducted in Cucurbitaceae plant breeding programs on the ripening process, phloem transport, disease resistance, cold tolerance and fruit quality traits. This paper presents the role played by new omic technologies in the creation of knowledge on the mechanisms of the formation of the breeding features. The analysis of NGS (NGS-next generation sequencing) data allows the discovery of new genes and regulatory sequences, their positions, and makes available large collections of molecular markers. Genome-wide expression studies provide breeders with an understanding of the molecular basis of complex traits. Firstly a high density map should be created for the reference genome, then each re-sequencing data could be mapped and new markers brought out into breeding populations. The paper also presents methods that could be used in the future for the creation of variability and genomic modification of the species in question. It has been shown also the state and usefulness in breeding the chloroplastomic and mitochondriomic study.

  1. Evaluating Variant Calling Tools for Non-Matched Next-Generation Sequencing Data

    Science.gov (United States)

    Sandmann, Sarah; de Graaf, Aniek O.; Karimi, Mohsen; van der Reijden, Bert A.; Hellström-Lindberg, Eva; Jansen, Joop H.; Dugas, Martin

    2017-02-01

    Valid variant calling results are crucial for the use of next-generation sequencing in clinical routine. However, there are numerous variant calling tools that usually differ in algorithms, filtering strategies, recommendations and thus, also in the output. We evaluated eight open-source tools regarding their ability to call single nucleotide variants and short indels with allelic frequencies as low as 1% in non-matched next-generation sequencing data: GATK HaplotypeCaller, Platypus, VarScan, LoFreq, FreeBayes, SNVer, SAMtools and VarDict. We analysed two real datasets from patients with myelodysplastic syndrome, covering 54 Illumina HiSeq samples and 111 Illumina NextSeq samples. Mutations were validated by re-sequencing on the same platform, on a different platform and expert based review. In addition we considered two simulated datasets with varying coverage and error profiles, covering 50 samples each. In all cases an identical target region consisting of 19 genes (42,322 bp) was analysed. Altogether, no tool succeeded in calling all mutations. High sensitivity was always accompanied by low precision. Influence of varying coverages- and background noise on variant calling was generally low. Taking everything into account, VarDict performed best. However, our results indicate that there is a need to improve reproducibility of the results in the context of multithreading.

  2. Molecular typing of lung adenocarcinoma on cytological samples using a multigene next generation sequencing panel.

    Directory of Open Access Journals (Sweden)

    Aldo Scarpa

    Full Text Available Identification of driver mutations in lung adenocarcinoma has led to development of targeted agents that are already approved for clinical use or are in clinical trials. Therefore, the number of biomarkers that will be needed to assess is expected to rapidly increase. This calls for the implementation of methods probing the mutational status of multiple genes for inoperable cases, for which limited cytological or bioptic material is available. Cytology specimens from 38 lung adenocarcinomas were subjected to the simultaneous assessment of 504 mutational hotspots of 22 lung cancer-associated genes using 10 nanograms of DNA and Ion Torrent PGM next-generation sequencing. Thirty-six cases were successfully sequenced (95%. In 24/36 cases (67% at least one mutated gene was observed, including EGFR, KRAS, PIK3CA, BRAF, TP53, PTEN, MET, SMAD4, FGFR3, STK11, MAP2K1. EGFR and KRAS mutations, respectively found in 6/36 (16% and 10/36 (28% cases, were mutually exclusive. Nine samples (25% showed concurrent alterations in different genes. The next-generation sequencing test used is superior to current standard methodologies, as it interrogates multiple genes and requires limited amounts of DNA. Its applicability to routine cytology samples might allow a significant increase in the fraction of lung cancer patients eligible for personalized therapy.

  3. Evaluating Variant Calling Tools for Non-Matched Next-Generation Sequencing Data

    Science.gov (United States)

    Sandmann, Sarah; de Graaf, Aniek O.; Karimi, Mohsen; van der Reijden, Bert A.; Hellström-Lindberg, Eva; Jansen, Joop H.; Dugas, Martin

    2017-01-01

    Valid variant calling results are crucial for the use of next-generation sequencing in clinical routine. However, there are numerous variant calling tools that usually differ in algorithms, filtering strategies, recommendations and thus, also in the output. We evaluated eight open-source tools regarding their ability to call single nucleotide variants and short indels with allelic frequencies as low as 1% in non-matched next-generation sequencing data: GATK HaplotypeCaller, Platypus, VarScan, LoFreq, FreeBayes, SNVer, SAMtools and VarDict. We analysed two real datasets from patients with myelodysplastic syndrome, covering 54 Illumina HiSeq samples and 111 Illumina NextSeq samples. Mutations were validated by re-sequencing on the same platform, on a different platform and expert based review. In addition we considered two simulated datasets with varying coverage and error profiles, covering 50 samples each. In all cases an identical target region consisting of 19 genes (42,322 bp) was analysed. Altogether, no tool succeeded in calling all mutations. High sensitivity was always accompanied by low precision. Influence of varying coverages- and background noise on variant calling was generally low. Taking everything into account, VarDict performed best. However, our results indicate that there is a need to improve reproducibility of the results in the context of multithreading. PMID:28233799

  4. Molecular Typing of Lung Adenocarcinoma on Cytological Samples Using a Multigene Next Generation Sequencing Panel

    Science.gov (United States)

    Fassan, Matteo; Rachiglio, Anna Maria; Cappellesso, Rocco; Antonello, Davide; Amato, Eliana; Mafficini, Andrea; Lambiase, Matilde; Esposito, Claudia; Bria, Emilio; Simonato, Francesca; Scardoni, Maria; Turri, Giona; Chilosi, Marco; Tortora, Giampaolo; Fassina, Ambrogio; Normanno, Nicola

    2013-01-01

    Identification of driver mutations in lung adenocarcinoma has led to development of targeted agents that are already approved for clinical use or are in clinical trials. Therefore, the number of biomarkers that will be needed to assess is expected to rapidly increase. This calls for the implementation of methods probing the mutational status of multiple genes for inoperable cases, for which limited cytological or bioptic material is available. Cytology specimens from 38 lung adenocarcinomas were subjected to the simultaneous assessment of 504 mutational hotspots of 22 lung cancer-associated genes using 10 nanograms of DNA and Ion Torrent PGM next-generation sequencing. Thirty-six cases were successfully sequenced (95%). In 24/36 cases (67%) at least one mutated gene was observed, including EGFR, KRAS, PIK3CA, BRAF, TP53, PTEN, MET, SMAD4, FGFR3, STK11, MAP2K1. EGFR and KRAS mutations, respectively found in 6/36 (16%) and 10/36 (28%) cases, were mutually exclusive. Nine samples (25%) showed concurrent alterations in different genes. The next-generation sequencing test used is superior to current standard methodologies, as it interrogates multiple genes and requires limited amounts of DNA. Its applicability to routine cytology samples might allow a significant increase in the fraction of lung cancer patients eligible for personalized therapy. PMID:24236184

  5. Pseudorandom Bit Sequence Generator for Stream Cipher Based on Elliptic Curves

    Directory of Open Access Journals (Sweden)

    Jilna Payingat

    2015-01-01

    Full Text Available This paper proposes a pseudorandom sequence generator for stream ciphers based on elliptic curves (EC. A detailed analysis of various EC based random number generators available in the literature is done and a new method is proposed such that it addresses the drawbacks of these schemes. Statistical analysis of the proposed method is carried out using the NIST (National Institute of Standards and Technology test suite and it is seen that the sequence exhibits good randomness properties. The linear complexity analysis shows that the system has a linear complexity equal to the period of the sequence which is highly desirable. The statistical complexity and security against known plain text attack are also analysed. A comparison of the proposed method with other EC based schemes is done in terms of throughput, periodicity, and security, and the proposed method outperforms the methods in the literature. For resource constrained applications where a highly secure key exchange is essential, the proposed method provides a good option for encryption by time sharing the point multiplication unit for EC based key exchange. The algorithm and architecture for implementation are developed in such a way that the hardware consumed in addition to point multiplication unit is much less.

  6. Unveiling Chloroplast RNA Editing Events Using Next Generation Small RNA Sequencing Data

    Directory of Open Access Journals (Sweden)

    Nureyev F. Rodrigues

    2017-09-01

    Full Text Available Organellar RNA editing involves the modification of nucleotide sequences to maintain conserved protein functions, mainly by reverting non-neutral codon mutations. The loss of plastid editing events, resulting from mutations in RNA editing factors or through stress interference, leads to developmental, physiological and photosynthetic alterations. Recently, next generation sequencing technology has generated the massive discovery of sRNA sequences and expanded the number of sRNA data. Here, we present a method to screen chloroplast RNA editing using public sRNA libraries from Arabidopsis, soybean and rice. We mapped the sRNAs against the nuclear, mitochondrial and plastid genomes to confirm predicted cytosine to uracil (C-to-U editing events and identify new editing sites in plastids. Among the predicted editing sites, 40.57, 34.78, and 25.31% were confirmed using sRNAs from Arabidopsis, soybean and rice, respectively. SNP analysis revealed 58.2, 43.9, and 37.5% new C-to-U changes in the respective species and identified known and new putative adenosine to inosine (A-to-I RNA editing in tRNAs. The present method and data reveal the potential of sRNA as a reliable source to identify new and confirm known editing sites.

  7. Detection of identity by descent using next-generation whole genome sequencing data

    Directory of Open Access Journals (Sweden)

    Su Shu-Yi

    2012-06-01

    Full Text Available Abstract Background Identity by descent (IBD has played a fundamental role in the discovery of genetic loci underlying human diseases. Both pedigree-based and population-based linkage analyses rely on estimating recent IBD, and evidence of ancient IBD can be used to detect population structure in genetic association studies. Various methods for detecting IBD, including those implemented in the soft- ware programs fastIBD and GERMLINE, have been developed in the past several years using population genotype data from microarray platforms. Now, next-generation DNA sequencing data is becoming increasingly available, enabling the comprehensive analysis of genomes, in- cluding identifying rare variants. These sequencing data may provide an opportunity to detect IBD with higher resolution than previously possible, potentially enabling the detection of disease causing loci that were previously undetectable with sparser genetic data. Results Here, we investigate how different levels of variant coverage in sequencing and microarray genotype data influences the resolution at which IBD can be detected. This includes microarray genotype data from the WTCCC study, denser genotype data from the HapMap Project, low coverage sequencing data from the 1000 Genomes Project, and deep coverage complete genome data from our own projects. With high power (78%, we can detect segments of length 0.4 cM or larger using fastIBD and GERMLINE in sequencing data. This compares to similar power to detect segments of length 1.0 cM or higher with microarray genotype data. We find that GERMLINE has slightly higher power than fastIBD for detecting IBD segments using sequencing data, but also has a much higher false positive rate. Conclusion We further quantify the effect of variant density, conditional on genetic map length, on the power to resolve IBD segments. These investigations into IBD resolution may help guide the design of future next generation sequencing studies that

  8. Coupled high-throughput functional screening and next generation sequencing for identification of plant polymer decomposing enzymes in metagenomic libraries

    OpenAIRE

    2013-01-01

    Recent advances in sequencing technologies generate new predictions and hypotheses about the functional roles of environmental microorganisms. Yet, until we can test these predictions at a scale that matches our ability to generate them, most of them will remain as hypotheses. Function-based mining of metagenomic libraries can provide direct linkages between genes, metabolic traits and microbial taxa and thus bridge this gap between sequence data generation and functional predictions. Here we...

  9. SRAdb: query and use public next-generation sequencing data from within R

    Directory of Open Access Journals (Sweden)

    Zhu Yuelin

    2013-01-01

    Full Text Available Abstract Background The Sequence Read Archive (SRA is the largest public repository of sequencing data from the next generation of sequencing platforms including Illumina (Genome Analyzer, HiSeq, MiSeq, .etc, Roche 454 GS System, Applied Biosystems SOLiD System, Helicos Heliscope, PacBio RS, and others. Results SRAdb is an attempt to make queries of the metadata associated with SRA submission, study, sample, experiment and run more robust and precise, and make access to sequencing data in the SRA easier. We have parsed all the SRA metadata into a SQLite database that is routinely updated and can be easily distributed. The SRAdb R/Bioconductor package then utilizes this SQLite database for querying and accessing metadata. Full text search functionality makes querying metadata very flexible and powerful. Fastq files associated with query results can be downloaded easily for local analysis. The package also includes an interface from R to a popular genome browser, the Integrated Genomics Viewer. Conclusions SRAdb Bioconductor package provides a convenient and integrated framework to query and access SRA metadata quickly and powerfully from within R.

  10. Application of next-generation sequencing in clinical oncology to advance personalized treatment of cancer

    Institute of Scientific and Technical Information of China (English)

    Yan-Fang Guan; Gai-Rui Li; Rong-Jiao Wang; Yu-Ting Yi; Ling Yang; Dan Jiang; Xiao-Ping Zhang; Yin Peng

    2012-01-01

    With the development and improvement of new sequencing technology,next-generation sequencing (NGS) has been applied increasingly in cancer genomics research over the past decade.More recently,NGS has been adopted in clinical oncology to advance personalized treatment of cancer.NGS is used to identify novel and rare cancer mutations,detect familial cancer mutation carriers,and provide molecular rationale for appropriate targeted therapy.Compared to traditional sequencing,NGS holds many advantages,such as the ability to fully sequence all types of mutations for a large number of genes (hundreds to thousands) in a single test at a relatively low cost.However,significant challenges,particularly with respect to the requirement for simpler assays,more flexible throughput,shorter turnaround time,and most importantly,easier data analysis and interpretation,will have to be overcome to translate NGS to the bedside of cancer patients.Overall,continuous dedication to apply NGS in clinical oncology practice will enable us to be one step closer to personalized medicine.

  11. Read length and repeat resolution: Exploring prokaryote genomes using next-generation sequencing technologies

    KAUST Repository

    Cahill, Matt J.

    2010-07-12

    Background: There are a growing number of next-generation sequencing technologies. At present, the most cost-effective options also produce the shortest reads. However, even for prokaryotes, there is uncertainty concerning the utility of these technologies for the de novo assembly of complete genomes. This reflects an expectation that short reads will be unable to resolve small, but presumably abundant, repeats. Methodology/Principal Findings: Using a simple model of repeat assembly, we develop and test a technique that, for any read length, can estimate the occurrence of unresolvable repeats in a genome, and thus predict the number of gaps that would need to be closed to produce a complete sequence. We apply this technique to 818 prokaryote genome sequences. This provides a quantitative assessment of the relative performance of various lengths. Notably, unpaired reads of only 150nt can reconstruct approximately 50% of the analysed genomes with fewer than 96 repeat-induced gaps. Nonetheless, there is considerable variation amongst prokaryotes. Some genomes can be assembled to near contiguity using very short reads while others require much longer reads. Conclusions: Given the diversity of prokaryote genomes, a sequencing strategy should be tailored to the organism under study. Our results will provide researchers with a practical resource to guide the selection of the appropriate read length. 2010 Cahill et al.

  12. Read length and repeat resolution: exploring prokaryote genomes using next-generation sequencing technologies.

    Directory of Open Access Journals (Sweden)

    Matt J Cahill

    Full Text Available BACKGROUND: There are a growing number of next-generation sequencing technologies. At present, the most cost-effective options also produce the shortest reads. However, even for prokaryotes, there is uncertainty concerning the utility of these technologies for the de novo assembly of complete genomes. This reflects an expectation that short reads will be unable to resolve small, but presumably abundant, repeats. METHODOLOGY/PRINCIPAL FINDINGS: Using a simple model of repeat assembly, we develop and test a technique that, for any read length, can estimate the occurrence of unresolvable repeats in a genome, and thus predict the number of gaps that would need to be closed to produce a complete sequence. We apply this technique to 818 prokaryote genome sequences. This provides a quantitative assessment of the relative performance of various lengths. Notably, unpaired reads of only 150nt can reconstruct approximately 50% of the analysed genomes with fewer than 96 repeat-induced gaps. Nonetheless, there is considerable variation amongst prokaryotes. Some genomes can be assembled to near contiguity using very short reads while others require much longer reads. CONCLUSIONS: Given the diversity of prokaryote genomes, a sequencing strategy should be tailored to the organism under study. Our results will provide researchers with a practical resource to guide the selection of the appropriate read length.

  13. Next-Generation Sequencing of Aquatic Oligochaetes: Comparison of Experimental Communities

    Science.gov (United States)

    Vivien, Régis; Lejzerowicz, Franck; Pawlowski, Jan

    2016-01-01

    Aquatic oligochaetes are a common group of freshwater benthic invertebrates known to be very sensitive to environmental changes and currently used as bioindicators in some countries. However, more extensive application of oligochaetes for assessing the ecological quality of sediments in watercourses and lakes would require overcoming the difficulties related to morphology-based identification of oligochaetes species. This study tested the Next-Generation Sequencing (NGS) of a standard cytochrome c oxydase I (COI) barcode as a tool for the rapid assessment of oligochaete diversity in environmental samples, based on mixed specimen samples. To know the composition of each sample we Sanger sequenced every specimen present in these samples. Our study showed that a large majority of OTUs (Operational Taxonomic Unit) could be detected by NGS analyses. We also observed congruence between the NGS and specimen abundance data for several but not all OTUs. Because the differences in sequence abundance data were consistent across samples, we exploited these variations to empirically design correction factors. We showed that such factors increased the congruence between the values of oligochaetes-based indices inferred from the NGS and the Sanger-sequenced specimen data. The validation of these correction factors by further experimental studies will be needed for the adaptation and use of NGS technology in biomonitoring studies based on oligochaete communities. PMID:26866802

  14. Evaluation of next generation sequencing for the analysis of Eimeria communities in wildlife.

    Science.gov (United States)

    Vermeulen, Elke T; Lott, Matthew J; Eldridge, Mark D B; Power, Michelle L

    2016-05-01

    Next-generation sequencing (NGS) techniques are well-established for studying bacterial communities but not yet for microbial eukaryotes. Parasite communities remain poorly studied, due in part to the lack of reliable and accessible molecular methods to analyse eukaryotic communities. We aimed to develop and evaluate a methodology to analyse communities of the protozoan parasite Eimeria from populations of the Australian marsupial Petrogale penicillata (brush-tailed rock-wallaby) using NGS. An oocyst purification method for small sample sizes and polymerase chain reaction (PCR) protocol for the 18S rRNA locus targeting Eimeria was developed and optimised prior to sequencing on the Illumina MiSeq platform. A data analysis approach was developed by modifying methods from bacterial metagenomics and utilising existing Eimeria sequences in GenBank. Operational taxonomic unit (OTU) assignment at a high similarity threshold (97%) was more accurate at assigning Eimeria contigs into Eimeria OTUs but at a lower threshold (95%) there was greater resolution between OTU consensus sequences. The assessment of two amplification PCR methods prior to Illumina MiSeq, single and nested PCR, determined that single PCR was more sensitive to Eimeria as more Eimeria OTUs were detected in single amplicons. We have developed a simple and cost-effective approach to a data analysis pipeline for community analysis of eukaryotic organisms using Eimeria communities as a model. The pipeline provides a basis for evaluation using other eukaryotic organisms and potential for diverse community analysis studies.

  15. Food Safety in the Age of Next Generation Sequencing, Bioinformatics, and Open Data Access.

    Science.gov (United States)

    Taboada, Eduardo N; Graham, Morag R; Carriço, João A; Van Domselaar, Gary

    2017-01-01

    Public health labs and food regulatory agencies globally are embracing whole genome sequencing (WGS) as a revolutionary new method that is positioned to replace numerous existing diagnostic and microbial typing technologies with a single new target: the microbial draft genome. The ability to cheaply generate large amounts of microbial genome sequence data, combined with emerging policies of food regulatory and public health institutions making their microbial sequences increasingly available and public, has served to open up the field to the general scientific community. This open data access policy shift has resulted in a proliferation of data being deposited into sequence repositories and of novel bioinformatics software designed to analyze these vast datasets. There also has been a more recent drive for improved data sharing to achieve more effective global surveillance, public health and food safety. Such developments have heightened the need for enhanced analytical systems in order to process and interpret this new type of data in a timely fashion. In this review we outline the emergence of genomics, bioinformatics and open data in the context of food safety. We also survey major efforts to translate genomics and bioinformatics technologies out of the research lab and into routine use in modern food safety labs. We conclude by discussing the challenges and opportunities that remain, including those expected to play a major role in the future of food safety science.

  16. Next-Generation Sequencing of Aquatic Oligochaetes: Comparison of Experimental Communities.

    Science.gov (United States)

    Vivien, Régis; Lejzerowicz, Franck; Pawlowski, Jan

    2016-01-01

    Aquatic oligochaetes are a common group of freshwater benthic invertebrates known to be very sensitive to environmental changes and currently used as bioindicators in some countries. However, more extensive application of oligochaetes for assessing the ecological quality of sediments in watercourses and lakes would require overcoming the difficulties related to morphology-based identification of oligochaetes species. This study tested the Next-Generation Sequencing (NGS) of a standard cytochrome c oxydase I (COI) barcode as a tool for the rapid assessment of oligochaete diversity in environmental samples, based on mixed specimen samples. To know the composition of each sample we Sanger sequenced every specimen present in these samples. Our study showed that a large majority of OTUs (Operational Taxonomic Unit) could be detected by NGS analyses. We also observed congruence between the NGS and specimen abundance data for several but not all OTUs. Because the differences in sequence abundance data were consistent across samples, we exploited these variations to empirically design correction factors. We showed that such factors increased the congruence between the values of oligochaetes-based indices inferred from the NGS and the Sanger-sequenced specimen data. The validation of these correction factors by further experimental studies will be needed for the adaptation and use of NGS technology in biomonitoring studies based on oligochaete communities.

  17. Next-Generation Sequencing of Aquatic Oligochaetes: Comparison of Experimental Communities.

    Directory of Open Access Journals (Sweden)

    Régis Vivien

    Full Text Available Aquatic oligochaetes are a common group of freshwater benthic invertebrates known to be very sensitive to environmental changes and currently used as bioindicators in some countries. However, more extensive application of oligochaetes for assessing the ecological quality of sediments in watercourses and lakes would require overcoming the difficulties related to morphology-based identification of oligochaetes species. This study tested the Next-Generation Sequencing (NGS of a standard cytochrome c oxydase I (COI barcode as a tool for the rapid assessment of oligochaete diversity in environmental samples, based on mixed specimen samples. To know the composition of each sample we Sanger sequenced every specimen present in these samples. Our study showed that a large majority of OTUs (Operational Taxonomic Unit could be detected by NGS analyses. We also observed congruence between the NGS and specimen abundance data for several but not all OTUs. Because the differences in sequence abundance data were consistent across samples, we exploited these variations to empirically design correction factors. We showed that such factors increased the congruence between the values of oligochaetes-based indices inferred from the NGS and the Sanger-sequenced specimen data. The validation of these correction factors by further experimental studies will be needed for the adaptation and use of NGS technology in biomonitoring studies based on oligochaete communities.

  18. Food Safety in the Age of Next Generation Sequencing, Bioinformatics, and Open Data Access

    Directory of Open Access Journals (Sweden)

    Eduardo N. Taboada

    2017-05-01

    Full Text Available Public health labs and food regulatory agencies globally are embracing whole genome sequencing (WGS as a revolutionary new method that is positioned to replace numerous existing diagnostic and microbial typing technologies with a single new target: the microbial draft genome. The ability to cheaply generate large amounts of microbial genome sequence data, combined with emerging policies of food regulatory and public health institutions making their microbial sequences increasingly available and public, has served to open up the field to the general scientific community. This open data access policy shift has resulted in a proliferation of data being deposited into sequence repositories and of novel bioinformatics software designed to analyze these vast datasets. There also has been a more recent drive for improved data sharing to achieve more effective global surveillance, public health and food safety. Such developments have heightened the need for enhanced analytical systems in order to process and interpret this new type of data in a timely fashion. In this review we outline the emergence of genomics, bioinformatics and open data in the context of food safety. We also survey major efforts to translate genomics and bioinformatics technologies out of the research lab and into routine use in modern food safety labs. We conclude by discussing the challenges and opportunities that remain, including those expected to play a major role in the future of food safety science.

  19. Personal microbiomes and next-generation sequencing for laboratory-based education.

    Science.gov (United States)

    Hartman, Mark R; Harrington, Kristin T; Etson, Candice M; Fierman, Matthew B; Slonim, Donna K; Walt, David R

    2016-12-01

    Sequencing and bioinformatics technologies have advanced rapidly in recent years, driven largely by developments in next-generation sequencing (NGS) technology. Given the increasing importance of these advances, there is a growing need to incorporate concepts and practices relating to NGS into undergraduate and high school science curricula. We believe that direct access to sequencing and bioinformatics will improve the ability of students to understand the information obtained through these increasingly ubiquitous research tools. In this commentary, we discuss approaches and challenges for bringing NGS into the classroom based on our experiences in developing and running a microbiome project in high school and undergraduate courses. We describe strategies for maximizing student engagement through establishing personal relevance and utilizing an inquiry-based structure. Additionally, we address the practical issues of incorporating cutting edge technologies into an established curriculum. Looking forward, we anticipate that NGS educational experiments will become more commonplace as sequencing costs continue to decrease and the workflow becomes more user friendly. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Relative Stability of Peptide Sequence Ions Generated by Tandem Mass Spectrometry

    Science.gov (United States)

    Bythell, Benjamin J.; Hendrickson, Christopher L.; Marshall, Alan G.

    2012-04-01

    We report the use of unimolecular dissociation by infrared radiation for gaseous multiphoton energy transfer to determine relative activation energy (Ea,laser) for dissociation of peptide sequence ions. The sequence ions of interest are mass-isolated; the entire ion cloud is then irradiated with a continuous wave CO2 laser, and the first order rate constant, kd, is determined for each of a series of laser powers. Provided these conditions are met, a plot of the natural logarithm of kd versus the natural logarithm of laser power yields a straight line, whose slope provides a measure of Ea,laser. This method reproduces the Ea values from blackbody radiative dissociation (BIRD) for the comparatively large, singly and doubly protonated bradykinin ions (nominally y 9 and y 9 2+ ). The comparatively small sequence ion systems produce Ea,laser values that are systematic underestimates of theoretical barriers calculated with density functional theory (DFT). However, the relative Ea,laser values are in qualitative agreement with the mobile proton model and available theory. Additionally, novel protonated cyclic-dipeptide (diketopiperazine) fragmentation reactions are analyzed with DFT. FT-ICR MS provides access to sequence ions generated by electron capture dissociation, infrared multiphoton dissociation, and collisional activation methods (i.e., b n , y m , c n , z m • ions).