Pfu

Mic Chaudoir mic at nwu.edu
Fri Jan 20 01:29:26 EST 1995


In article <D_henrique-1601950951180001 at mac030005.ox.icnet.uk>,
D_henrique at icrf.icnet.uk (Domingos henrique) wrote:

> Dear netters
> Can I have the experienced oppinion of someone that uses routinely Pfu
> polymerase for amplifyng products for cloning. Is it worth to prevent
> mistakes in the PCR? I want to use the PCR product for cloning and then
> expression in E.coli and am afraid of mistakes that can result in
> truncated proteins.
> Thanks
> Domingos

Our lab does NOT routinely use pfu, but we have used it for some
site-directed mutagenesis.  We had bad experiences:

1)  We still had errors!!  The error rate seemed to be about the same as
normal taq.  Our cDNA was only 500 bp, so this was not a huge deal.  Juts
had to sequence a lot of clones.

2)  Pfu conditions are different than taq, so you have to work out
conditions again.

I would reccomend trying NEB vent or something...

-- 
mic
mic at nwu.edu
 
"It [PowerPC Mac] won't have any effect at all on Intel machines.
They will continue to plod along, running the same clunky Windows              and wretched DOS drivel they always have.  The PPC will affect Intel PCs the same way an SR-71 Blackbird affects a dairy cow."
                     Robert Rhode, in comp.sys.mac.advocacy



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