Book ChapterDOI
Real-time DNA sequencing from single polymerase molecules.
Jonas Korlach,Keith Bjornson,Bidhan Chaudhuri,Ronald L. Cicero,Benjamin Flusberg,Jeremy Gray,David P. Holden,Ravi Saxena,Jeffrey Wegener,Stephen Turner +9 more
TLDR
This chapter will first outline the principle of this single-molecule, real-time (SMRT) DNA sequencing method, followed by descriptions of its underlying components and typical sequencing run conditions.Abstract:
Pacific Biosciences has developed a method for real-time sequencing of single DNA molecules (Eid et al., 2009), with intrinsic sequencing rates of several bases per second and read lengths into the kilobase range. Conceptually, this sequencing approach is based on eavesdropping on the activity of DNA polymerase carrying out template-directed DNA polymerization. Performed in a highly parallel operational mode, sequential base additions catalyzed by each polymerase are detected with terminal phosphate-linked, fluorescence-labeled nucleotides. This chapter will first outline the principle of this single-molecule, real-time (SMRT) DNA sequencing method, followed by descriptions of its underlying components and typical sequencing run conditions. Two examples are provided which illustrate that, in addition to the DNA sequence, the dynamics of DNA polymerization from each enzyme molecules is directly accessible: the determination of base-specific kinetic parameters from single-molecule sequencing reads, and the characterization of DNA synthesis rate heterogeneities.read more
Citations
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疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI
Sequencing technologies-the next generation
TL;DR: A technical review of template preparation, sequencing and imaging, genome alignment and assembly approaches, and recent advances in current and near-term commercially available NGS instruments is presented.
Journal ArticleDOI
De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis
Brian J. Haas,Alexie Papanicolaou,Moran Yassour,Moran Yassour,Manfred Grabherr,Philip D. Blood,Joshua C. Bowden,M. B. Couger,David Eccles,Bo Li,Matthias Lieber,Matthew D. MacManes,Michael Ott,Joshua Orvis,Nathalie Pochet,Nathalie Pochet,Francesco Strozzi,Nathan T. Weeks,Rick Westerman,Thomas William,Colin N. Dewey,Robert Henschel,Richard D. LeDuc,Nir Friedman,Aviv Regev +24 more
TL;DR: This protocol provides a workflow for genome-independent transcriptome analysis leveraging the Trinity platform and presents Trinity-supported companion utilities for downstream applications, including RSEM for transcript abundance estimation, R/Bioconductor packages for identifying differentially expressed transcripts across samples and approaches to identify protein-coding genes.
Journal ArticleDOI
Nonhybrid, finished microbial genome assemblies from long-read SMRT sequencing data
Chen-Shan Chin,David Alexander,Patrick Marks,Aaron Klammer,James P Drake,Cheryl Heiner,Alicia Clum,Alex Copeland,John Huddleston,Evan E. Eichler,Stephen Turner,Jonas Korlach +11 more
TL;DR: This work presents a hierarchical genome-assembly process (HGAP) for high-quality de novo microbial genome assemblies using only a single, long-insert shotgun DNA library in conjunction with Single Molecule, Real-Time (SMRT) DNA sequencing.
Journal ArticleDOI
A tale of three next generation sequencing platforms: comparison of Ion Torrent, Pacific Biosciences and Illumina MiSeq sequencers.
Michael A. Quail,Miriam Smith,Paul Coupland,Thomas D. Otto,Simon R. Harris,Thomas R. Connor,Anna Bertoni,Harold Swerdlow,Yong Gu +8 more
TL;DR: 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.
References
More filters
疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A
TL;DR: PfPMP1)与感染红细胞、树突状组胞以及胎盘的单个或多个受体作用,在黏附及免疫逃避中起关键的作�ly.
Journal ArticleDOI
Real-Time DNA Sequencing from Single Polymerase Molecules
John Eid,Adrian Fehr,Jeremy Gray,Khai Luong,John Lyle,Geoff Otto,Paul Peluso,David R. Rank,Primo Baybayan,Brad Bettman,Arkadiusz Bibillo,Keith Bjornson,Bidhan Chaudhuri,Fred Christians,Ronald L. Cicero,Sonya Clark,Ravindra V. Dalal,Alex DeWinter,John Dixon,Mathieu Foquet,Alfred Gaertner,Paul Hardenbol,Cheryl Heiner,Kevin Hester,David P. Holden,Gregory J. Kearns,Xiangxu Kong,Ronald Kuse,Yves Lacroix,Steven Lin,Paul Lundquist,Congcong Ma,Patrick Marks,Mark Maxham,Devon Murphy,Insil Park,Thang Pham,Michael Phillips,Joy Roy,Robert Sebra,Gene Shen,Jon M. Sorenson,Austin B. Tomaney,Kevin Travers,Mark Trulson,John Vieceli,Jeffrey Wegener,Dawn Wu,Alicia Yang,Denis Zaccarin,Peter Zhao,Frank Zhong,Jonas Korlach,Stephen Turner +53 more
TL;DR: Single-molecule, real-time sequencing data obtained from a DNA polymerase performing uninterrupted template-directed synthesis using four distinguishable fluorescently labeled deoxyribonucleoside triphosphates (dNTPs) are presented.
Journal ArticleDOI
Next-Generation DNA Sequencing Methods
TL;DR: An astounding potential exists for next-generation DNA sequencing technologies to bring enormous change in genetic and biological research and to enhance the authors' fundamental biological knowledge.
Journal ArticleDOI
Shotgun bisulphite sequencing of the Arabidopsis genome reveals DNA methylation patterning
Shawn J. Cokus,Suhua Feng,Xiaoyu Zhang,Zugen Chen,Barry Merriman,Christian D. Haudenschild,Sriharsa Pradhan,Stanley F. Nelson,Matteo Pellegrini,Steven E. Jacobsen +9 more
TL;DR: A map at single-base-pair resolution of methylated cytosines for Arabidopsis is generated by combining bisulphite treatment of genomic DNA with ultra-high-throughput sequencing using the Illumina 1G Genome Analyser and Solexa sequencing technology.
Journal ArticleDOI
An optimal extraction algorithm for ccd spectroscopy.
TL;DR: In this article, an optimal spectrum extraction procedure is described, and examples of its performance with CCD data are presented, which delivers the maximum possible signal-to-noise ratio while preserving spectrophotometric accuracy.
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