PfuTurbo (hotstart version)

Frederik Wirtz-Peitz fw218 at cam.ac.uk
Tue Nov 13 14:10:25 EST 2001


The enzyme yields about 5X as much product in Pwo buffer compared to the
native buffer. What a joke.

But that is still only a small fraction of Pwo's yield.

Frederik


"Frederik Wirtz-Peitz" <fw218 at cam.ac.uk> wrote in message
news:9sr2uo$eu7$1 at pegasus.csx.cam.ac.uk...
> Extension time with Pwo was 1 min/kb. PfuTurbo fails at both 1 min/kb and
2
> min/kb. Actually, 2 min/kb gave me a faint band of about 10 ng (I ran out
> approx. 10% of the reaction). Otherwise the program was identical, yes.
>
> Maybe the Pwo buffer results in decreased fidelity. But what good is a
> buffer that gives me the best fidelity but no product?
>
> Anyway, I will post the results of Pfu in Pwo buffer later.
>
> The Pwo isn't hotstart but still didn't give any unspecific product. I
don't
> understand myself. Why am I wasting all this time and money?
>
> Thanks so far,
>
> Frederik
>
>
> "Roland Hübner" <roland.hubner at ua.ac.be> wrote in message
> news:3bf103c2$1 at news.uia.ac.be...
> > > If Pwo and Pfu are identical, then the buffer must be responsible for
> any
> > > discrepancies.
> >
> > Hello,
> >
> >  of course the buffers are different AND affect Hifi of the pol...
> > Perhaps, and there is certainly some lit about that, more processivity =
> > lower fidelity?
> >
> > Cheers,
> > Roland
> >
> >
> >  BTW, was the cycling prg. (especially ext. time) identical in your
> > comparisons?
>
>





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