cloning of PCR products - errors?

Ian A. York iayork at panix.com
Wed Jun 27 09:01:27 EST 2001


In article <3B39CD96.B8AB041 at bbm1.ucm.es>,
Sergio  <sergioal at bbm1.ucm.es> wrote:
>
>Only 265x10-6 ??... damn... i must be doing something wrong. We use to get
>around 1x10-3 mutations when using Taq... almost five times the supposed error
>rate.

I think the published error rates are for *optimized* PCR with the various 
enzymes.  Mixing Primer A and Primer B and throwing it in with template 
isn't optimizing the reaction.  I suspect that if you really fiddled with 
the conditions you could get a much higher fidelity.

Ian 
-- 
    Ian York   (iayork at panix.com)  <http://www.panix.com/~iayork/>
    "-but as he was a York, I am rather inclined to suppose him a
     very respectable Man." -Jane Austen, The History of England




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