poor cloning efficiency w/Pfu turbo

Duncan Clark junk at hgmp.mrc.ac.uk
Wed Jul 31 05:49:00 EST 2002


Historians believe that in newspost <ai7u8a$91t$1 at netnews.upenn.edu> on 
Wed, 31 Jul 2002, Paul Shinn <pshinn at mail2.sas.upenn.edu> penned the 
following literary masterpiece:
> After PCR, they are sodium acetate
>precipitated and then digested with Sfi overnight.

Taq survives Ethanol pptation and some nucleotides also ppt.

Pfu may survive even better.

You then land up with a nice gap filling enzyme during your overnight 
Sfi I digest so your overhangs get blunted hence lower cloning 
efficiency.

The survivability is documented by Wayne Barnes in Gene several years 
ago in his Blue/white fidelity assay for KlenTaq.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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