Problem using pfuturbo for PCR

Thomas Burmeister tburmei at zedat.fu-berlin.de
Fri Jun 18 16:01:58 EST 1999


You could also try to use phosphorothioate oligonucleotides (see Skerra,
Nucleic Acids Res. 1992 Jul 25;20(14):3551-4) to eliminate this possible
primer shortening.

Thomas Burmeister

In article <F6QJvQAG9Ka3Ew3y at genesys.demon.co.uk>, "Dr. Duncan Clark"
<Duncan at genesys.demon.co.uk> wrote:

>I wonder if the 3'-5' exonuclease activity of the PfuTurbo is shortening
>your primers (in the time it takes to prepare the PCR mix and get it to
>94C) such that they then anneal elsewhere on your target. Try adding the
>enzyme to the PCR mix in tubes at 94C on the PCR machine i.e. a manual
>hotstart.

-- 
Thomas Burmeister
<mailto:tbu at gmx.net>



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