Pfu polymerase

Ian A. York iayork at panix.com
Tue Dec 24 11:47:29 EST 1996


In article <59p0eu$5tr at news.jhu.edu>,
PETER S. SILVERSTEIN <silverps at welchlink.welch.jhu.edu> wrote:
>Does anyone out there have any first hand experience with Pfu polymerase 
>from Stratagene?  If so, how does it work?  Is it better than Vent?  I 
>have been having some problems with Vent and have been thinking of switching.

I haven't compared it to Vent; but I've used Pfu several times with no
real problems.  I use it when I'm PCRing for subsequent cloning, and use
Taq for 'diagnostic' PCR.  Taq definitely gives more DNA, but the error
rate of Pfu is much, much lower (somewhere on the web there's a comparison
of the various polymerases' error rates; Pfu is either the best or near
the best, as I recall).  I've been able to get my fragment each time with
the Pfu, but have had to do more fiddling with conditions than I usually
have with Taq; on the other hand, because I'm cloning with the Pfu my
primers are usually suboptimal because my start and/or end point is
fixed, so this isn't really a fair comparison.

I've used vent a couple of times and it took about the same amount of
fiddling to get my clone, so I don't know if there's much to recommend one
over the other.

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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