RT high GC% content template

yang gyang at MED.UNC.EDU
Thu Sep 23 16:06:37 EST 1999


Dear all,

I have some trouble for reverse transcription of high GC% content RNA (
800 bp, GC%=76% ).

Although I try to RT @ 65 degrees using ThermoSript RT system from BRL,
no PCR bands at all!    However, with other high GC% template ,  I can
get the right size PCR product by adding DMSO into my reaction buffer .
So, the problem with my RT-PCR lies in RT step.  Checking from gel, My
RNA is good before and after DNase I  treatment.
Probably RT step contribute to my failure?

Does anyone has some tricks in RT such hard RNA ?  Any suggestions will
be greatly appreciated.

Yang
UNC-CH, USA




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