Pfu/Taq in PCR reactions

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Thu Mar 2 15:31:33 EST 1995


On 26 Feb 1995 jnewton at molbiol.ox.ac.uk wrote:
 
> ...I've seen a number of 
> people advocate mixing Taq DNA polymerase with Pfu DNA polymerase in order to 
> improve/reduce the rate of mismatched base incorperation during PCR, however, 
> I can't understand why this should help.
 
...and on 28 Feb 1995 segdoh at uxa.cso.uiuc.edu wrote: 
 
| The reason some PCR kits have Pfu and Taq mixed together id to reduce the
| error rate (Pfu) and to facilitate TA cloning (Taq).  Pfu does not leave
| nontemplate terminal A overhangs like Taq which are required for TA cloning.
 
Hold on! Now I'm confused. Is it better for TA cloning to have a mixture of
polymerases including Pfu which will polish the A's off the 3' end, or
NOT to have Pfu ...allowing Taq to add the A's to the 3' end of the PCR product?
 
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