In article <8742i0$khm$1 at willow.cc.kcl.ac.uk>, Colin Dolphin
<colin.dolphin at kcl.ac.uk> writes
>It appears to me
>that the "best enzyme" might be a highly processive enzyme (most standard
>Taqs) in combination with a dash of Pfu, or equivalent, to add
>proof-reading activity together with a hot-start facility achieved,
>perhaps, by adding appropriate antibodies.
I would not call Taq highly processive! It has very poor processivity
(40-60bp) but is better by a factor of 2-4 than Pfu etc. Don't forget
that all enzymes currently used for PCR are repair enzymes not
replication enzymes.
Does Pfu need to be hot-started? Stratagene in a former Strategies
claimed that Pfu had minimal activity at RT so there shouldn't be a
problem with it.
So try a 'Gold' Taq with Pfu or an ordinary Taq with antibody and Pfu.
Duncan
--
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sliced from time to time....
Duncan Clark
DNAmp Ltd.
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