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Chunk #1 — Background

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Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2.
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Besides the need to account for the specifics of count data, such as non-normality and a dependence of the variance on the mean, a core challenge is the small number of samples in typical HTS experiments – often as few as two or three replicates per condition. Inferential methods that treat each gene separately suffer here from lack of power, due to the high uncertainty of within-group variance estimates. In high-throughput assays, this limitation can be overcome by pooling information across genes, specifically, by exploiting assumptions about the similarity of the variances of different genes measured in the same experiment [1].