Hi all, I have a primary question about preparing RNA samples. We have cancer tissue. We extract the RNA from the cell, then make cDNA from the RNA. We fragment the cDNA and size select. Then we send them for RNA-seq?
If is it like this, when a gene is expressed more, we will have more RNA strands associated to it in our sample. So we would have more reads for that RNA strand(gene) and so on....
So it is possible that we will have more duplicates for a highly expressed gene. If during the analysis we omit those duplicates we will loose information which shows we have had more expression related to that gene?
Thanks a lot in advance Narges