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Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2

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TLDR
This work presents DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates, which enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression.
Abstract
In comparative high-throughput sequencing assays, a fundamental task is the analysis of count data, such as read counts per gene in RNA-seq, for evidence of systematic changes across experimental conditions. Small replicate numbers, discreteness, large dynamic range and the presence of outliers require a suitable statistical approach. We present DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates. This enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression. The DESeq2 package is available at http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html .

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Citations
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Journal ArticleDOI

HTSeq—a Python framework to work with high-throughput sequencing data

TL;DR: This work presents HTSeq, a Python library to facilitate the rapid development of custom scripts for high-throughput sequencing data analysis, and presents htseq-count, a tool developed with HTSequ that preprocesses RNA-Seq data for differential expression analysis by counting the overlap of reads with genes.
Journal ArticleDOI

Comprehensive Integration of Single-Cell Data.

TL;DR: A strategy to "anchor" diverse datasets together, enabling us to integrate single-cell measurements not only across scRNA-seq technologies, but also across different modalities.
Journal ArticleDOI

Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown

TL;DR: This protocol describes all the steps necessary to process a large set of raw sequencing reads and create lists of gene transcripts, expression levels, and differentially expressed genes and transcripts.
References
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Journal ArticleDOI

Classification and clustering of sequencing data using a Poisson model

TL;DR: In this paper, the authors propose new approaches for performing classification and clustering of observations on the basis of sequencing data using a Poisson log linear model, which is an analog of diagonal linear discriminant analysis.
Journal ArticleDOI

Errors in RNA-Seq quantification affect genes of relevance to human disease

TL;DR: It is shown that it is possible to use data that may otherwise have been discarded to measure group-level expression, and that such data contains biologically relevant information.
Journal ArticleDOI

A powerful and flexible approach to the analysis of RNA sequence count data

TL;DR: BBSeq is described, which incorporates a simple beta-binomial generalized linear model, combined with simple outlier detection and testing approaches, which appears to have favorable characteristics in power and flexibility.
Posted ContentDOI

Salmon: Accurate, Versatile and Ultrafast Quantification from RNA-seq Data using Lightweight-Alignment

TL;DR: Salmon is introduced, a novel method and software tool for transcript quantication that exhibits state-of-the-art accuracy while being signicantly faster than most other tools.
Journal ArticleDOI

Classification and clustering of sequencing data using a Poisson model

Daniela Witten
- 28 Feb 2012 - 
TL;DR: Using a Poisson log linear model, an analog of diagonal linear discriminant analysis that is appropriate for sequencing data is developed and an approach for clustering sequencing data using a new dissimilarity measure that is based upon the Poisson model is proposed.
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