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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

Comparison of Affymetrix GeneChip expression measures

TL;DR: It is found that background correction, one of the main steps in preprocessing, has the largest effect on performance and, in particular, background correction appears to improve accuracy but, in general, worsen precision.
Journal ArticleDOI

Robustly detecting differential expression in RNA sequencing data using observation weights

TL;DR: The robustness of existing approaches for count-based differential expression analysis are studied and a new strategy based on observation weights that can be used within existing frameworks are proposed and the results suggest that outliers can have a global effect on differential analyses.
Posted Content

Robustly detecting differential expression in RNA sequencing data using observation weights

TL;DR: In this article, the authors study the robustness of existing approaches for count-based differential expression analysis and propose a new strategy based on observation weights that can be used within existing frameworks.
Journal ArticleDOI

Conservation of an RNA Regulatory Map Between Drosophila and Mammals

TL;DR: The RNA regulatory map of Pasilla (PS) and NOVA1/2 is highly conserved between insects and mammals despite the fact that the target gene orthologs regulated by PS and NoVA1 /2 are almost entirely nonoverlapping.
Journal ArticleDOI

Evaluating Gene Expression in C57BL/6J and DBA/2J Mouse Striatum Using RNA-Seq and Microarrays

TL;DR: This study compares RNA-Seq (Illumina GA IIx) with two microarray platforms to detect differential striatal gene expression between the B6 and D2 inbred mouse strains and shows that by using stringent data processing requirements differential expression is concordant with both the Affymetrix and Illumina platforms in more instances than it is concORDant with only a single platform.
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