PfuTurbo (hotstart version)
fw218 at cam.ac.uk
Thu Nov 15 08:00:22 EST 2001
Turbo or not, the polymerase remains identical. And in my case this magical
cofactor kills processivity, it does not enhance it. Simply a money-making
scheme if you ask me.
"Duncan Clark" <d_clark at genesys.demon.co.uk> wrote in message
> In message <9sqv9m$bdt$1 at pegasus.csx.cam.ac.uk>, Frederik Wirtz-Peitz
> <fw218 at cam.ac.uk> writes on the Tue, 13 Nov 2001
> >However, in my hands Pwo works and PfuTurbo doesn't. Pfu only fails in
> >reactions primed by long oligos (60mers in my case). The same template
> >amplified by shorter primers works fine. I do not understand why this is
> >case. I hear from someone else using Pfu that PAGE purification helps.
> >Again, it eludes me. Even if bases are missing from the 5' end, why would
> >this affect polymerization?
> >If Pwo and Pfu are identical, then the buffer must be responsible for any
> >discrepancies. Pwo buffer is just straight MgSO4, while Pfu's is more
> >complex and has loads of detergent in it.
> Pfu is identical to Pwo but PfuTurbo is an entirely different beast.
> PfuTurbo is Pfu plus to protein polymerase enhancing factors (some very
> very long US patent app last year). These basically act to enhance the
> 'long' PCR ability of Pfu whilst retaining the fidelity of Pfu.
> It explains why some batches of native enzyme were way better than the
> recombinant enzyme, as the native batches were 'contaminated' by these
> two extra proteins.
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
> Duncan Clark
> GeneSys Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
More information about the Methods