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Updated: Jul 6, 2025

Three Differential Expression Analysis Methods for RNA Sequencing: limma, EdgeR, DESeq2
Published on: September 18, 2021
Enhanced clustering-based differential expression analysis method for RNA-seq data
Manon Makino1, Kentaro Shimizu1, Koji Kadota1,2,3
1Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Abstract:
RNA-seq is a tool for measuring gene expression and is commonly used to identify differentially expressed genes (DEGs). Gene clustering has been widely used to classify DEGs with similar expression patterns, but rarely used to identify DEGs themselves. We recently reported that the clustering-based method (called MBCdeg1 and 2) for identifying DEGs has great potential. However, these methods left room for improvement. This study reports on the improvement (named MBCdeg3). We compared a total of six competing methods: three conventional R packages (edgeR, DESeq2, and TCC) and three versions of MBCdeg (i.e., MBCdeg1, 2, and 3) corresponding to three different normalization algorithms. As MBCdeg3 performs well in many simulation scenarios of RNA-seq count data, MBCdeg3 replaces MBCdeg1 and 2 in our previous report. •MBCdeg3 is a method for both identification and classification of DEGs from RNA-seq count data.•MBCdeg3 is available as a function of R, which is common in the field of expression analysis.•MBCdeg3 performs well in a variety of simulation scenarios for RNA-seq count data.

