Chatomics Field GuideWhat They Don't Teach You

Topic

Log transformation of counts

Should I use log2(x+1), vst or rlog, and why does the pseudocount matter?

Log of zero is undefined, so everyone adds a pseudocount, and the choice of pseudocount changes fold changes for low-expressed genes more than the biology does. Variance-stabilizing transforms exist precisely to avoid this, and no log-transformed matrix should ever be fed into a count-based test.