I am running gene set analysis on curated gene sets from the MSigDB. I was successfully able to run roast using the following coding
library("edgeR") summarized.counts <- read.table("C:/Users/Alicia/Documents/summarized.counts.matrix", row.names=1, header=TRUE, sep="\t") design_two<-model.matrix(~0+ factor(c(1,1,1,2,2,2,3,3,3))) colnames(design_two)<-c("stage1", "stage2", "stage3") con<-makeContrasts(stage3-stage1, levels=design_two) library(org.Hs.eg.db) gene_set<-c("CYSLTR2", "GPR17","LTB4R", "LTB4R2", "GNB5", "GIP", "GNB2", "SCT", "VIP", "GNG8") ind<-ids2indices(gene_set, row.names(summarized.counts)) ind dge.edgeR=DGEList(counts=summarized.counts) dge.edgeR<-calcNormFactors(dge.edgeR) dge.edgeR=estimateDisp(dge.edgeR, design_two, robust=TRUE) rst<-mroast(dge.edgeR, index=ind, design=design_two, nrot=9999, contrast=con) rst
I am concerned about my results though.
NGenes PropDown PropUp Direction PValue FDR PValue.Mixed FDR.Mixed
Set1 3 0.3333333 0.6666667 Up 0.0626 0.0626 1e-04 1e-04
Why would the output indicate a NGene number of 3 when I included 10 genes in my set?
(this is just practice data, my final gene sets will contain larger quantities of gene"
Thanks, I figured that was the issue was I'm not super confident with this stuff yet. I really appreciate your help and that command line you provided.