Hi all, while looking for SNPs for virus quasispecies in a badnavirus using RNA that was gel-excised with maximum length of 100, maybe 200 bp I find definitive read alignment hotspots to the master genome in all 3 samples. generally people have been finding these hotspots in the populations of siRNA of 21,22 or 24 nt length, products of silencing. however the hotspots I find are in the slightly longer reads. Sequencing ran for 36 cycles, so they are 36 nt long. Also, 90+% of these reads with potential lengths between 30 and 100 nt are aligning to the sense strand. I am trying to come up with an explanation for these two things and drawing a blank. I am not a virologist or even microbiologist, so am I missing something obvious? I apologize if this is not the place to ask this question, and would appreciate any answers or other places to ask this question.
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