Entering edit mode
3.5 years ago
chatterjee.arumoy
▴
50
Hi,
If I have done my RNA seq with 4 samples X 3 replicates each in two batches (see below for the design matrix):
1) Is it correct to adjust batch effects on the raw count file first by ComBat-seq (Zhang et.al., 2020) and then process the output by edgeR?
CONDITIONS SAMPLES BATCH
T1 R1 1
T1 R2 1
T1 R3 1
T2 R1 1
T2 R2 2
T2 R3 2
T3 R1 1
T3 R2 1
T3 R3 1
T4 R1 1
T4 R2 2
T4 R3 2
I am asking this question since the only R2 and R3 for conditions T2 and T4 were processed in batch 2 and I could not include such a batch effect in the edgeR model matrix (see below) which needs batch 2 to be present in each of T1, T2, T3 and T4.
design = model.matrix(~ 0 + conditions + batch, data=y$samples)
Ref: Zhang et.al., 2020: https://academic.oup.com/nargab/article/2/3/lqaa078/5909519
Kindly help.
Thanks
Arumoy.