(by shifalich from rediffmail.com)
Thu Sep 10 08:58:37 EST 2009
I also don't do RNaseH treatment, I was told by company guys that RNA being an unstable molecule will get degraded by itself in the second step. Also, you can check the RT that you are using is RNase H + or negative.
Hope this helps!!!!!!!
On Wed, 09 Sep 2009 21:02:11 +0530 wrote
I am doing reverse transcription with Superscript II and oligo(dT)s as the first step of a 2-step RT qPCR. I will do relative quantification on fragments not exceeding 250 bp in size.
How necessary is it to RNAseH treat my cDNA/RNA after reverse transcription? If I don't, will it cause trouble downstream?
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