Working on fine tuning mRNA isolation out of CTCs from blood. Tackling RNA degradation during the lysis and isolation step and we are considering RNase inhibitors to use in our reagents. Does anyone have recommendations on RNase inhibitors that won't interfere with downstream sequencing and qPCR? I have used RNase Out before but that method didn't include RNAseq.
Thank you!
I add RNase OUT when i'm performing lysis then move onto Trizol and use Zymo's direct-zol kit for isolation with on-column DNase digestion. This also removes the RNase OUT from the sample. just make sure once you are lysed and start the isolation procedure that you are careful with handling the tubes and are mindful of incubation times.
If you are running out of time then prep the lysate and freeze at -80 and isolate on another day
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