Pfu/Taq in PCR reactions

hodges bradley l segdoh at uxa.cso.uiuc.edu
Fri Mar 3 13:55:38 EST 1995


It is better to have just TAq for TA cloning.  Pfu added to the mix will 
increase fidelity of replication and also allows the amplification of 
longer templates.  I don't think Pfu will cleave off terminal A 
overhangs.  Or if it does the effect on TA cloning may be minimal.  So if 
you are cloning PCR products less than 1.5Kb, TAq in your mix will be 
sufficient.

On Thu, 2 Mar 1995, Paul N Hengen wrote:

> 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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