Hello, I am working on RNASEQ data with 4 treatments(Control, ABA, salt,sorbitol), 3 genotypes(col.0, lbd16-1, ds2.3) and three replicates. To test differently expressed genes of treated/untreated effect between lbd16 and col.0, I wrote my script like this:
dds <- DESeqDataSetFromMatrix(countdata, colData, design= ~ genotype + treatment + genotype:treatment)
dds$treatment <- relevel(dds$treatment , "Control")
dds$genotype <- relevel(dds$genotype , "Col.0")
keep <- rowSums(counts(dds)) >= 1
dds_F <- dds[keep,]
dds2 <- DESeq(dds_F)
dds2$group <- factor(paste0(dds2$genotype, dds2$treatment))
design(dds2) <- ~ group
resultsNames(dds2)
[1] "Intercept" "genotype_ds2.3_vs_Col.0" "genotype_lbd16.1_vs_Col.0"
[4] "treatment_ABA_vs_Control" "treatment_NaCl_vs_Control" "treatment_sorbitol_vs_Control"
[7] "genotypeds2.3.treatmentABA" "genotypelbd16.1.treatmentABA" "genotypeds2.3.treatmentNaCl"
[10] "genotypelbd16.1.treatmentNaCl" "genotypeds2.3.treatmentsorbitol" "genotypelbd16.1.treatmentsorbitol".
My question is:
- Is it the right script to find DEGs among treaments and genotypes?
- will
res<-results(dds2, contrast=list( c("treatment_ABA_vs_Control","genotypeds2.3.treatmentABA")))
give me the result that differently expressed genes of ABAvscontrol between ds2.3 and col.0. If not, what would be the right one? - If I just want to see DEGs of ABAvscontrol of Col0, is this right?
results(dds, contrast=c("treatment","ABA","Control"))
- If I just want to see DEGs of ABAvscontrol of lbd16, is this right?
results(dds, name="genotypelbd16.1.treatmentABA")
Thanks for your reply. I am not sure if I understand correctly.
So if aiming to answer treatment effect (e.g ABA treatment) between(not separately) genotype ds2.3 and reference genotype, using results(dds, name="genotypeds2.3.treatmentABA") is the right way. For treatment effect in a non-reference genotype, results(dds, list( c("treatmentABAvs_Control","genotypeds2.3.treatmentABA") )) will answer it.