Non-templated nucleotide addition in PCR

Martin Kennedy cytogen at chmeds.ac.nz
Mon Oct 26 17:58:42 EST 1992


 In article <1992Oct26.113558.17435 at syma.sussex.ac.uk>, bapr7 at syma.sussex.ac.uk (Jansen Jacob) writes:
> I am making site directed mutagenesis using PCR and am worried about
> non-templated nucleotide addition at the 3' end by TAQ polymerase. 
> Has anyone got any experience with Vent polymerase or any other
> polymerase which does not have this non-templated nucleotide addition.
> 
> 			Jansen Jacob
> 			AFRC Institute of Plant Science Research
> 			Nitrogen Fixation Laboratory
> 			University of Sussex
> 			Brighton BN1 9Q
> 			U.K.
> 
>

I've cloned VENT PCR products directly into Bluescript without any
pre-treatment of the fragments, and it seems to work fine.  Admittedly, the
fragments were entirely composed of G and C, and I used only dCTP and dGTP in
the reaction.  These VENT products certainly clone better than ordinary Taq Pol
derived products that have been trimmed with T4 pol.

Cheers,

Martin



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