Publication Date

3-15-2022

Journal

Genome Biology

DOI

10.1186/s13059-022-02648-4

PMID

35292087

PMCID

PMC8922736

PubMedCentral® Posted Date

3-15-2022

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Computational Biology, Gene Expression Profiling, Humans, RNA-Seq, Sample Size, Sequence Analysis, RNA

Abstract

When identifying differentially expressed genes between two conditions using human population RNA-seq samples, we found a phenomenon by permutation analysis: two popular bioinformatics methods, DESeq2 and edgeR, have unexpectedly high false discovery rates. Expanding the analysis to limma-voom, NOISeq, dearseq, and Wilcoxon rank-sum test, we found that FDR control is often failed except for the Wilcoxon rank-sum test. Particularly, the actual FDRs of DESeq2 and edgeR sometimes exceed 20% when the target FDR is 5%. Based on these results, for population-level RNA-seq studies with large sample sizes, we recommend the Wilcoxon rank-sum test.

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