Pfu Polymerase Questions
cooperj at nhlbi.nih.gov
Tue Aug 31 11:15:31 EST 1993
In article <1993Aug31.150238.576 at Virginia.EDU> mgk2r at Virginia.EDU (Michael G. Kurilla) writes:
>From: mgk2r at Virginia.EDU (Michael G. Kurilla)
>Subject: Re: Pfu Polymerase Questions
>Message-ID: <1993Aug31.150238.576 at Virginia.EDU>
>Organization: University of Virginia
>References: <1993Aug30.210453.15569 at fcom.cc.utah.edu>
>Date: Tue, 31 Aug 1993 15:02:38 GMT
>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
Where can I get a the Lander Program....
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