Does RNA - PCR ??

rmoldwin at midway.uchicago.edu rmoldwin at midway.uchicago.edu
Tue Jul 18 17:32:48 EST 1995


In article <aquilla.1156467038E at sadye.emba.uvm.edu>, aquilla at salus.med.uvm.edu 
(Tracy Aquilla) writes:

>In Article <3ue8ub$cam at saba.info.ucla.edu>, walter
><wlech at medicine.medsch.ucla.edu> wrote:
>>My understanding is that the RNA would be degraded rather rapidly under 
>>the usual PCR conditions via magnesium-catalyzed hydrolysis.
>
>Actually, RNA amplifies quite nicely with Taq polymerase. I have cloned
>several cDNAs from total RNA this way. It works.
>    tracy

I also have seen several reports of RT activity in Taq.  Perhaps, it varies 
according to the manufacturer, and the specific origin of the Taq, e.g. 
recombinant/modified vs. wild type.  In my lab, we very rarely see this RT 
activity when using standard Perkin Elmer recombinant AmpliTaq, which PE 
reports in their catalog to have "minimal, low" RT activity.  AmpliTaq is 
modified from the wild-type Taq, but I can't determine what the difference is 
from their literature.

I wouldn't not be surprised if changing PCR conditions could increase the RT 
activity.

So I guess you could try a different source of Taq, in addition to the previous 
suggestions.

Hope this helps,

--Rich




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