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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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Book

In all likelihood : statistical modelling and inference using likelihood

Yudi Pawitan
TL;DR: This paper presents a meta-modelling framework for estimating the likelihood of random parameters in a discrete-time environment and describes its use in simple and complex models.
Journal ArticleDOI

EBSeq: an empirical Bayes hierarchical model for inference in RNA-seq experiments.

TL;DR: EBSeq is developed, using the merits of empirical Bayesian methods, for identifying DE isoforms in an RNA-seq experiment comparing two or more biological conditions and proves to be a robust approach for identifying De genes.
Journal ArticleDOI

Small-sample estimation of negative binomial dispersion, with applications to SAGE data

TL;DR: A quantile-adjusted conditional maximum likelihood estimator for the dispersion parameter of the negative binomial distribution is derived and an "exact" test is derived that outperforms the standard approximate asymptotic tests.
Journal ArticleDOI

Multiple-laboratory comparison of microarray platforms

TL;DR: A consortium of ten laboratories from the Washington, DC–Baltimore, USA, area was formed to compare data obtained from three widely used platforms using identical RNA samples to demonstrate that there are relatively large differences in data obtained in labs using the same platform, but that the results from the best-performing labs agree rather well.
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