Optimized murine sample sizes for RNA sequencing studies revealed from large scale comparative analysis.

ABSTRACT: Determining the appropriate sample size (N) for bulk RNA sequencing experiments is critical for obtaining reliable results. We show in two N = 30 profiling studies, comparing wild-type mice and mice in which one copy of a gene has been deleted, the N required to minimize false positives and maximize true discoveries found in the N = 30 experiment. Results from experiments with N = 4 or less are shown to be highly misleading, given the high false positive rate and the lack of discovery of genes later found with higher N. For a cut-off of 2-fold expression differences, we find an N of 6-7 mice is required to consistently decrease the false positive rate to below 50%, and the detection sensitivity to above 50%. More is always better for both metrics - and an N of 8-12 is significantly better

SUBMITTER: Halasz G 

PROVIDER: S-EPMC12630869 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature