Pfu polymerase mixed with Taq polymerase

Austin P. So (Hae-Jin) nobody at nowhere.com
Wed May 24 14:14:49 EST 2006


Julio Cortazar wrote:

> Have heared that one may mix Pfu and Taq polymerases (Fermantas). Taq
> is copying Pfu verifying. Have any of u tried this combbination? I'm
> planning to introduce two nucleotide changes in 1kb DNA fragment. So
> far I made set of 6 PCRs (I'm using so called SOE-PCR method with
> overlapping extensions) but my RedTaq polymerase made so many mistakes
> that I started to sworn it very badly.

I remember looking into this, and if my memory serves me correct, a 1:9 
mixture of Pfu to Taq is used for analysis of single nucleotide 
polymorphisms by an HPLC based technique. I don't know if anyone has 
tried it for longer templates though.

That being said, I don't remember ever finding a unit concentration that 
people used, or what people used to dilute the polymerase mixtures.

Austin

P.S. another thing you may consider is getting your overlapping primers 
(?) purified...at least find out what the purity of your primers are.


More information about the Methods mailing list