Journal ArticleDOI
Sequencing depth and coverage: key considerations in genomic analyses
TLDR
The issue of sequencing depth in the design of next-generation sequencing experiments is discussed and current guidelines and precedents on the issue of coverage are reviewed for four major study designs, including de novo genome sequencing, genome resequencing, transcriptome sequencing and genomic location analyses.Abstract:
Sequencing technologies have placed a wide range of genomic analyses within the capabilities of many laboratories. However, sequencing costs often set limits to the amount of sequences that can be generated and, consequently, the biological outcomes that can be achieved from an experimental design. In this Review, we discuss the issue of sequencing depth in the design of next-generation sequencing experiments. We review current guidelines and precedents on the issue of coverage, as well as their underlying considerations, for four major study designs, which include de novo genome sequencing, genome resequencing, transcriptome sequencing and genomic location analyses (for example, chromatin immunoprecipitation followed by sequencing (ChIP-seq) and chromosome conformation capture (3C)).read more
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A survey of best practices for RNA-seq data analysis
Ana Conesa,Pedro Madrigal,Pedro Madrigal,Sonia Tarazona,David Gomez-Cabrero,Alejandra Cervera,Andrew McPherson,Michał Wojciech Szcześniak,Daniel J. Gaffney,Laura L. Elo,Xuegong Zhang,Ali Mortazavi +11 more
TL;DR: All of the major steps in RNA-seq data analysis are reviewed, including experimental design, quality control, read alignment, quantification of gene and transcript levels, visualization, differential gene expression, alternative splicing, functional analysis, gene fusion detection and eQTL mapping.
Journal ArticleDOI
Estimating the population abundance of tissue-infiltrating immune and stromal cell populations using gene expression
Etienne Becht,Nicolas A. Giraldo,Nicolas A. Giraldo,Nicolas A. Giraldo,Laetitia Lacroix,Laetitia Lacroix,Laetitia Lacroix,Bénédicte Buttard,Bénédicte Buttard,Bénédicte Buttard,Nabila Elarouci,Florent Petitprez,Janick Selves,Pierre Laurent-Puig,Catherine Sautès-Fridman,Catherine Sautès-Fridman,Catherine Sautès-Fridman,Wolf H. Fridman,Wolf H. Fridman,Wolf H. Fridman,Aurélien de Reyniès +20 more
TL;DR: The Microenvironment Cell Populations-counter method is introduced, which allows the robust quantification of the absolute abundance of eight immune and two stromal cell populations in heterogeneous tissues from transcriptomic data and demonstrates that MCP-counter overcomes several limitations or weaknesses of previously proposed computational approaches.
Journal ArticleDOI
Recovery of nearly 8,000 metagenome-assembled genomes substantially expands the tree of life
Donovan H. Parks,Christian Rinke,Maria Chuvochina,Pierre-Alain Chaumeil,Ben J. Woodcroft,Paul N. Evans,Philip Hugenholtz,Gene W. Tyson +7 more
TL;DR: The recovery of 7,903 bacterial and archaeal metagenome-assembled genomes increases the phylogenetic diversity represented by public genome repositories and provides the first representatives from 20 candidate phyla.
Journal ArticleDOI
Qualimap 2: advanced multi-sample quality control for high-throughput sequencing data
TL;DR: Qualimap 2 represents a next step in the QC analysis of HTS data, along with comprehensive single-sample analysis of alignment data, and includes new modes that allow simultaneous processing and comparison of multiple samples.
Journal ArticleDOI
UMI-tools: modeling sequencing errors in Unique Molecular Identifiers to improve quantification accuracy
TL;DR: It is shown that errors in the UMI sequence are common and network-based methods to account for these errors when identifying PCR duplicates are introduced, demonstrating the value of properly accounting for errors in UMIs.
References
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The Transcriptional Landscape of the Yeast Genome Defined by RNA Sequencing
Ugrappa Nagalakshmi,Zhong Wang,Karl Waern,C. Shou,Debasish Raha,Mark Gerstein,Michael Snyder +6 more
TL;DR: A quantitative sequencing-based method is developed for mapping transcribed regions, in which complementary DNA fragments are subjected to high-throughput sequencing and mapped to the genome, and it is demonstrated that most (74.5%) of the nonrepetitive sequence of the yeast genome is transcribed.
Journal ArticleDOI
The ENCODE (ENCyclopedia of DNA elements) Project
Elise A. Feingold,Peter J. Good,Mark S. Guyer,S. Kamholz,L. Liefer,Kris A. Wetterstrand,Francis S. Collins,Thomas R. Gingeras,Dione Kampa,E. A. Sekinger,Jill Cheng,H. Hirsch,Srinka Ghosh,Zhou Zhu,Sandeep Patel,Antonio Piccolboni,A. Yang,Hari Tammana,Stefan Bekiranov,Philipp Kapranov,Rhonda Harrison,George M. Church,Kevin Struhl,Bing Ren,Tae Hoon Kim,Leah O. Barrera,Chunxu Qu,S. van Calcar,R. Luna,C. K. Glass,M. G. Rosenfeld,Roderic Guigó,Stylianos E. Antonarakis,Ewan Birney,Michael R. Brent,Lior Pachter,Alexandre Reymond,Emmanouil T. Dermitzakis,Colin N. Dewey,Damian Keefe,F. Denoeud,Julien Lagarde,Jennifer L. Ashurst,Tim Hubbard,Jan-Jaap Wesselink,Robert Castelo,Eduardo Eyras,Richard M. Myers,Arend Sidow,Serafim Batzoglou,Nathan D. Trinklein,Sara J. Hartman,Shelley Force Aldred,Elizabeth Anton,Diane I. Schroeder,S. S. Marticke,Long H. Nguyen,Jeremy Schmutz,Jane Grimwood,Mark Dickson,Gregory M. Cooper,Eric A. Stone,George Asimenos,Michael Brudno,Anindya Dutta,Neerja Karnani,Christopher M. Taylor,H. K. Kim,Gabriel Robins,George Stamatoyannopoulos,John A. Stamatoyannopoulos,Michael O. Dorschner,Peter J. Sabo,Michael Hawrylycz,Richard Humbert,James C. Wallace,Min-Feng Yu,Patrick A. Navas,M. McArthur,William Stafford Noble,Ian Dunham,Christof Koch,Robert M. Andrews,Gayle K. Clelland,Sarah Wilcox,Joanna C. Fowler,Keith D. James,P. Groth,Oliver M. Dovey,Peter D. Ellis,Vicki L. Wraight,Andrew J. Mungall,Pawandeep Dhami,Heike Fiegler,Cordelia Langford,Nigel P. Carter,David Vetrie,Michael Snyder,Ghia Euskirchen,Alexander E. Urban,Ugrappa Nagalakshmi,John L. Rinn,George V. Popescu,Paul Bertone,Stephen Hartman,Joel Rozowsky,Olof Emanuelsson,Thomas Royce,Sambath Chung,Mark Gerstein,Zheng Lian,Jane B. Lian,Y. Nakayama,Sherman M. Weissman,Viktor Stolc,W. Tongprasit,H. Sethi,Steven J.M. Jones,Marco A. Marra,H. Shin,Jacquie Schein,Michele Clamp,Kerstin Lindblad-Toh,Jean L. Chang,David B. Jaffe,Michael Kamal,Eric S. Lander,Tarjei S. Mikkelsen,Jade P. Vinson,Michael C. Zody,P. J. De Jong,Kazutoyo Osoegawa,Mikhail Nefedov,Baoli Zhu,Andreas D. Baxevanis,Tyra G. Wolfsberg,Gregory E. Crawford,James R R Whittle,Ingeborg Holt,T. J. Vasicek,D. Zhou,Shu-Jin Luo,Eric D. Green,Gerry Bouffard,Elliott H. Margulies,Matthew E. Portnoy,Nancy F. Hansen,Pamela J. Thomas,Jenny McDowell,Baishali Maskeri,Alice C. Young,Jacquelyn R. Idol,Robert W. Blakesley,Greg Schuler,Webb Miller,Ross C. Hardison,Laura Elnitski,Prachi Shah,Steven L. Salzberg,Mihaela Pertea,William H. Majoros,David Haussler,Daryl J. Thomas,Kate R. Rosenbloom,Hiram Clawson,Adam Siepel,W. J. Kent,Zhiping Weng,S. Jin,Anason S. Halees,H. Burden,Ulas Karaoz,Yutao Fu,Yong Yu,C. Ding,Charles R. Cantor,Robert E. Kingston,Jonathan H. Dennis,Roland Green,Michael A. Singer,Todd Richmond,J. E. Norton,Peggy J. Farnham,Matthew J. Oberley,D. R. Inman,M. R. McCormick,H. Kim,C. L. Middle,Michael C. Pirrung,Xiang-Dong Fu,Y. S. Kwon,Z. Ye,Job Dekker,Tomoko M. Tabuchi,Nele Gheldof,Josée Dostie,S. C. Harvey +196 more
TL;DR: The ENCyclopedia Of DNA Elements (ENCODE) Project is organized as an international consortium of computational and laboratory-based scientists working to develop and apply high-throughput approaches for detecting all sequence elements that confer biological function.
Journal ArticleDOI
ChIP-Seq: advantages and challenges of a maturing technology
TL;DR: The benefits and challenges in harnessing ChIP–seq are described with an emphasis on issues related to experimental design and data analysis, and effective computational analysis will be crucial for uncovering biological mechanisms.
Journal ArticleDOI
ChIP-seq guidelines and practices of the ENCODE and modENCODE consortia
Stephen G. Landt,Georgi K. Marinov,Anshul Kundaje,Pouya Kheradpour,Florencia Pauli,Serafim Batzoglou,Bradley E. Bernstein,Peter J. Bickel,James B. Brown,Philip Cayting,Yiwen Chen,Gilberto DeSalvo,Charles B. Epstein,Katherine I. Fisher-Aylor,Ghia Euskirchen,Mark Gerstein,Jason Gertz,Alexander J. Hartemink,Michael M. Hoffman,Vishwanath R. Iyer,Youngsook L. Jung,Subhradip Karmakar,Manolis Kellis,Peter V. Kharchenko,Qiang Li,Tao Liu,X. Shirley Liu,Lijia Ma,Aleksandar Milosavljevic,Richard M. Myers,Peter J. Park,Michael J. Pazin,Marc D. Perry,Debasish Raha,Timothy E. Reddy,Joel Rozowsky,Noam Shoresh,Arend Sidow,Matthew Slattery,John A. Stamatoyannopoulos,Michael Y. Tolstorukov,Kevin P. White,Simon Xi,Peggy J. Farnham,Jason D. Lieb,Barbara J. Wold,Michael Snyder +46 more
TL;DR: This work discusses how ChIP quality, assessed in these ways, affects different uses of ChIP-seq data and develops a set of working standards and guidelines for ChIP experiments that are updated routinely.
Journal ArticleDOI
Evaluation of statistical methods for normalization and differential expression in mRNA-Seq experiments
TL;DR: This work provides a detailed evaluation of statistical methods for normalization and differential expression analysis of Illumina transcriptome sequencing (mRNA-Seq) data and investigates the impact of the read count normalization method on DE results, and proposes more general quantile-based normalization procedures.