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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David R. Bentley,Shankar Balasubramanian,Harold Swerdlow,Harold Swerdlow,Geoffrey Paul Smith,John Milton,John Milton,Clive Gavin Brown,Clive Gavin Brown,Kevin Hall,Dirk J. Evers,Colin Barnes,Colin Barnes,Helen Bignell,Jonathan Mark Boutell,Jason Bryant,Richard J. Carter,R. Keira Cheetham,Anthony J. Cox,Darren James Ellis,Michael R. Flatbush,Niall Anthony Gormley,Sean Humphray,Leslie J. Irving,Mirian S. Karbelashvili,Scott M. Kirk,Heng Li,Xiaohai Liu,Xiaohai Liu,Klaus Maisinger,Lisa Murray,Bojan Obradovic,Tobias William Barr Ost,Michael Lawrence Parkinson,M. R. Pratt,Isabelle Rasolonjatovo,Mark T. Reed,Roberto Rigatti,Chiara Rodighiero,Mark T. Ross,Andrea Sabot,Subramanian V. Sankar,Aylwyn Scally,Gary P. Schroth,Mark Smith,Vincent Peter Smith,Anastassia Spiridou,Peta E. Torrance,Svilen S. Tzonev,Eric Vermaas,Klaudia Walter,Wu Xiaolin,Lu Zhang,Mohammed D. Alam,Carole Anastasi,Ify C. Aniebo,David Mark Dunstan Bailey,Iain R. Bancarz,Saibal Banerjee,Selena G. Barbour,Primo Baybayan,Vincent A. Benoit,Kevin Benson,Claire Bevis,Phillip J. Black,Asha Boodhun,Joe S. Brennan,John Bridgham,Rob C. Brown,Andrew A. Brown,Dale Buermann,Abass A. Bundu,James C. Burrows,Nigel P. Carter,Nestor Castillo,Maria Chiara E. Catenazzi,Simon Chang,R. Neil Cooley,Natasha R. Crake,Olubunmi O. Dada,Konstantinos D. Diakoumakos,Belen Dominguez-Fernandez,David James Earnshaw,David James Earnshaw,Ugonna C. Egbujor,David W. Elmore,Sergey Etchin,Mark R. Ewan,Milan Fedurco,Louise Fraser,Karin Fuentes Fajardo,W. Scott Furey,David George,Kimberley J. Gietzen,Colin P. Goddard,George Stefan Golda,Philip A. Granieri,David E. Green,David L. Gustafson,Nancy F. Hansen,Kevin Harnish,Christian D. Haudenschild,Narinder I. Heyer,Matthew M. Hims,Johnny T. Ho,Adrian Horgan,Katya Hoschler,Steve Hurwitz,Denis V. Ivanov,Maria Q. Johnson,Terena James,T. A. Huw Jones,Gyoung-Dong Kang,Tzvetana H. Kerelska,Alan D. Kersey,Irina Khrebtukova,Alex P. Kindwall,Zoya Kingsbury,Paula Kokko-Gonzales,Anil Kumar,Marc Laurent,Cindy Lawley,Sarah E. Lee,Xavier Lee,Arnold Liao,Jennifer A. Loch,Mitch Lok,Shujun Luo,Radhika M. Mammen,John W. Martin,Patrick Mccauley,Paul McNitt,Parul Mehta,Keith W. Moon,Joe W. Mullens,Taksina Newington,Zemin Ning,Bee Ling Ng,Sonia M. Novo,Michael J. O'Neill,Mark A. Osborne,Mark A. Osborne,Andrew Osnowski,Omead Ostadan,Lambros L. Paraschos,Lea Pickering,Andrew C. Pike,Alger C. Pike,D. Chris Pinkard,Daniel P. Pliskin,Joe Podhasky,Victor J. Quijano,Come Raczy,Vicki H. Rae,Stephen Rawlings,Ana Chiva Rodriguez,Phyllida M. Roe,John Rogers,Maria Candelaria Rogert Bacigalupo,Nikolai Romanov,Anthony Romieu,Rithy K. Roth,Natalie J. Rourke,Silke Ruediger,Eli Rusman,Raquel Maria Sanches-Kuiper,Martin R. Schenker,Josefina M. Seoane,Richard Shaw,Mitch K. Shiver,Steven W. Short,Ning Sizto,Johannes P. Sluis,Melanie Anne Smith,Jean Ernest Sohna Sohna,Eric J. Spence,Kim B. Stevens,Neil Sutton,Lukasz Szajkowski,Carolyn Tregidgo,Gerardo Turcatti,Stephanie Vandevondele,Yuli Verhovsky,Selene M. Virk,Suzanne Wakelin,Gregory C. Walcott,Jingwen Wang,Graham John Worsley,Juying Yan,Ling Yau,Mike Zuerlein,Jane Rogers,James C. Mullikin,Matthew E. Hurles,Nick J. McCooke,Nick J. McCooke,John Stephen West,Frank L. Oaks,Peter Lundberg,David Klenerman,Richard Durbin,Anthony J. Smith +201 more
TL;DR: An approach that generates several billion bases of accurate nucleotide sequence per experiment at low cost is reported, effective for accurate, rapid and economical whole-genome re-sequencing and many other biomedical applications.
Journal ArticleDOI
Differential analysis of gene regulation at transcript resolution with RNA-seq
Cole Trapnell,David G. Hendrickson,David G. Hendrickson,Martin Sauvageau,Martin Sauvageau,Loyal A. Goff,Loyal A. Goff,John L. Rinn,John L. Rinn,Lior Pachter +9 more
TL;DR: Cuffdiff 2, an algorithm that estimates expression at transcript-level resolution and controls for variability evident across replicate libraries, robustly identifies differentially expressed transcripts and genes and reveals differential splicing and promoter-preference changes.
Journal ArticleDOI
Integrative annotation of human large intergenic noncoding RNAs reveals global properties and specific subclasses
Moran N. Cabili,Cole Trapnell,Cole Trapnell,Loyal A. Goff,Magdalena J. Koziol,Magdalena J. Koziol,Barbara Tazon-Vega,Barbara Tazon-Vega,Aviv Regev,John L. Rinn,John L. Rinn +10 more
TL;DR: It is found that lincRNA expression is strikingly tissue-specific compared with coding genes, and that l incRNAs are typically coexpressed with their neighboring genes, albeit to an extent similar to that of pairs of neighboring protein-coding genes.
Journal ArticleDOI
Genome-Wide Mapping of in Vivo Protein-DNA Interactions
David S. Johnson,Ali Mortazavi,Ali Mortazavi,Richard M. Myers,Richard M. Myers,Barbara J. Wold,Barbara J. Wold +6 more
TL;DR: A large-scale chromatin immunoprecipitation assay based on direct ultrahigh-throughput DNA sequencing was developed, which was then used to map in vivo binding of the neuron-restrictive silencer factor (NRSF; also known as REST) to 1946 locations in the human genome.
Journal ArticleDOI
Transcriptome-wide Identification of RNA-Binding Protein and MicroRNA Target Sites by PAR-CLIP
Markus Hafner,Markus Landthaler,Lukas Burger,Mohsen Khorshid,Jean Hausser,Philipp Berninger,Andrea Rothballer,Manuel Ascano,Anna-Carina Jungkamp,Mathias Munschauer,Alexander Ulrich,Greg S. Wardle,Scott Dewell,Mihaela Zavolan,Thomas Tuschl +14 more
TL;DR: This study developed a cell-based crosslinking approach to determine at high resolution and transcriptome-wide the binding sites of cellular RBPs and miRNPs and revealed that these factors bind thousands of sites containing defined sequence motifs and have distinct preferences for exonic versus intronic or coding versus untranslated transcript regions.