I want to normalize gene expression values per sample .

In doing this, TMM normalization is necessary.

My data is WTS data which is consist of raw count.

And there are 32 samples. Some of them are response group and the others are non-response group.

My question :

1. I think TMM normalization isn't related to group. Is it right?

2. my cod for TMM normalization like this:

   library(tweeDEseq)

   gp<-rep(1,the number of groups)
   
   F.norm<-normalizeCounts(df,group=gp)
   TMM<-F.norm+1
   

if there are any problems, correct my code If other good method for TMM normalization, please let me know