I got totally confused when looking at edgeR manual somehow:
The are different ways of estimating the dispersions (GLM and a classic approach). Also, one might estimate either CommonDisp or TrendedDisp.
I get totally different results when applying GLM and classic and also when excluding TrendedDisp.
My experiment is quite simple: I have 2 treated and 2 untreated samples and want to find differential expressed genes.
I also found that I have a batch effect, so that I got batch-values from RUVseq. What would be an applicable method (GLM or classic) to use if I want to intergrate batch effect into the design matrix. Also, when should I use estimateGLMTrendedDisp over estimateGLMCommonDisp?
GLM:
y <- estimateGLMCommonDisp(y, design) y <- estimateGLMTrendedDisp(y, design) y <- estimateGLMTagwiseDisp(y, design) then apply glmFit and glmLRT
classic: y <- estimateCommonDisp(y) y <- estimateTagwiseDisp(y) then apply exact test
EDIT:
also I get different BCV plots when I run either y <- estimateDisp(y, design) or those three commands together
y <- estimateGLMCommonDisp(y, design) y <- estimateGLMTrendedDisp(y, design) y <- estimateGLMTagwiseDisp(y, design)
what is quite weird because according to the edger manual they should give the same results.
Define "totally different". Totally different dispersions? Totally different DE genes?
I get different DE genes and different BCV plots and also strange p.values. With the "trended dispersion" most of the p.values are 1.