Historians believe that in newspost <[email protected]> on
Tue, 17 Aug 2010, DK <[email protected]> penned the following
literary masterpiece:
I would like to do everything humanly possible to reduce the
frequency of PCR errors in some reactions. I use PfuUltra
Fusion, which has probably highest fidelity of them all.
Pfu with their modified Sso7d domain plus Pfu pol- enzyme to give extra
proof-reading exo activity. Also dUTPase as well?
It also is very processive. I am wondering if reducing
an extension temperature (say, to 65C) will reduce not
only processivity
I would expect the processivity to stay the same regardless of
temperature and just the extension rate to be slower. Although dropping
from 72 to 65 will have negligible effect on the rate.
but also an error rate. Or will the error
rate remain the same?
Again I would expect to stay the same.
I guess the answer depend on the precise mechanism
of polymerization errors (e.g., which are the ones that
escape proofreading corrections?) - and I just know know
what it is.
Maybe Wayne Barnes or one the Stratagene people have published what the
standard error format is for Pfu
Aha.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC146123/.
Other than keeping the cycles as low as possible and lots of initial
template there's not too much more you can do.
Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
Duncan Clark
GeneSys Ltd.
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