RT-PCR of highly coiled RNA
vgrubor at pgrad.unimelb.edu.au
Tue Jan 29 04:01:51 EST 2002
Have you tried using Tth Polymerase? It can be obtained from Promega among
others and it has reverse transcriptase activity at 70-75 (can't remember
the exact number.
Roland Hübner wrote:
>> I have been trying to amplify by RT-PCR a region with very high secondary
>> structure. In order to reduce this I have tried to rise the temperature,
>> DMSO, change the KCl concentration, and add glicerol and even formamide.
>> Nothings seems to work! Any suggestion?
> what was the "highest" temperature you tried so far ?
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