Hi, I am using 26 .CEL files having 3 disease conditions from GEO for DGE analysis. The PCA plot after RMA normalization shows mixing of samples together.
So when I use
BATCH.cor <- readData(NORM.data, factor = PHENO)
BATCH.data <- ARSyNseq(BATCH.cor, factor="Group", batch = FALSE, norm = "n", logtransf = TRUE)
to remove batch effect it separates the samples on PCA .
But when is use
varFilter(BATCH.data, var.func=sd, var.cutoff=0.25, filterByQuantile=TRUE)
it reduces the total number of genes from 54000 to 10000.
Also i get more number of DOWN DEG than UP DEG after performing eBayes.
Is there any other simple batch correction and gene filter package available that didn't interfere with results ?
I answered almost the same question from you 3 days ago: DEG Filtering. It is a bit disappointing to find that you haven't taken any notice of my advice. I told you that the varFilter and ARSyNSeq steps were wrong yet you have continued them without any change, Should I conclude that you don't want any advice from me in the future?