Hi All, I am interested in identifying bacterial genes that are differentially expressed in vivo in comparison to the in vitro bacterial culture. However, I encountered a challenge regarding the sequencing depth of my samples. For in vivo bacterial RNA samples(total 8 samples) the sequencing depth was low in the range of 250K-1.3 million reads due to the presence of a large amount of host mRNA. For in vitro bacterial cultures (3 samples) the total reads were in the range of 70-100 million. My question is can DESeq2 handle such huge variations in sequencing depth among the samples? If not, can I down-sample the in vitro bacterial reads perhaps around 10-20 million may be using seqtk tool? I greatly appreciate any inputs you can provide on this matter. Thanks, Gopi Viswanathan
Thanks for your suggestion, Michael. I will try that strategy.