RT-PCR of highly coiled RNA

Vladimir Grubor 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,
>> add
>> DMSO, change the KCl concentration, and add glicerol and even formamide.
>> Nothings seems to work! Any suggestion?
> 
> Hi,
> 
>  what was the "highest" temperature you tried so far ?
> 
> R.




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