noncomplementary primers amd pfu polymerase

Duncan Clark junk at hgmp.mrc.ac.uk
Mon Feb 4 14:20:17 EST 2002


Historians believe that in 
<001f01c1ad68$8d9c0160$9293a8c0 at uz.kuleuven.ac.be> on the Mon, 4 Feb 
2002 that Peter Cherepanov <peter.cherepanov at uz.kuleuven.ac.be> penned 
the following literary masterpiece
>After few failures with Pfu, I switched to Pwo polymerase in all my cloning
>work; it is more processive then Pfu, always works better (makes a bit more
>mistakes though, but if you do not cycle the reaction too many times and
>take care to sequence your construct at the end, it is not going to be a
>problem).

This does not make much scientific sense.

Pfu and Pwo have the same DNA sequence (check Genbank or EMBL), 
therefore the same amino acid sequence and there should be no difference 
between the recombinant enzymes.

Any difference in performance should therefore be down to say reaction 
buffers or maybe the purification procedure on the enzymes.

Duncan

-- 
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....

Duncan Clark
GeneSys Ltd.




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