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Pfu Polymerase Questions

Michael G. Kurilla mgk2r at Virginia.EDU
Tue Aug 31 10:02:38 EST 1993


Stillman Lab  writes:
> 	I am using a recombinant PCR technique to create transcriptional fusions
> between two homologous genes. I have recovered several clones which
> contain Taq induced mutations, and I am trying to find a better enzyme to
> perform my PCR reactions. 
> 	Stratagene markets the Pfu Polymerase which is supposed to have a much
> lower error rate than either Taq or Vent. Has anyone actually used Pfu
> and compared the rate himself? If someone has used Pfu in PCR could you
> tell me if the extension time and/or temperature varies from those for
> Taq. Thanks in advance for your reply.
>  
> Helen_McBride at hlthsci.med.utah.edu

We have used Pfu for cloning experiments.  After several
constructs and sequencing, we have found about 1 clone in 10
has a mutation (not bad I think).  Generally, the sizes have
ranged from 300 - 1100 bases.

The Pfu salt conditions are different than Taq (10mM vs 50mM).
that corresponds to a reduction in the melting temperature by
about 10 - 11 degrees.  We use the Lander Primer program which
allows input of salt concentration so it hasn't been a problem
for us.

Michael Kurilla



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