PfuTurbo (hotstart version)
d_clark at genesys.demon.co.uk
Thu Nov 15 05:18:58 EST 2001
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 the
>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....
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