Pfu

Aaron Pawlyk aaron at netnews.upenn.edu
Sun Jan 22 10:11:51 EST 1995


Duncan Clark (Duncan at genesys.demon.co.uk) wrote:
: In article: <3fmojq$bjn at netnews.upenn.edu>  aaron at netnews.upenn.edu (Aaron Pawlyk) writes:

: > What I and several others have tried with great success is to use Taq and Pfu
: > in a ratio of 100 to 1.  (eg. 1 unit of Pfu for 100 units of Taq).  This allows
: > Taq to perform the bulk of the polymerase work with Pfu serving to check for
: > errors and avoids and excess exonuclease activity.  I know people who have 
: > checked the error rate with these and they are on the order to 1 per 10 million
: > base pairs.

: 1 in 10,000,000. I don't believe it. From Wayne Barnes published work with
: KlenTaq/Pfu the experimental error rate is nowhere near this. If you get 1 
: in 20,000 you are doing very well. I agree that the mix is very good but
: can we at least be realistic.

: Duncan  

: -----------------------------------------------------------------------------
: My mind's made up. Don't confuse me with the facts!

Ooops.  Misread my notes.  That should be 1 in 100,000 (according to the 
notes).  I never checked the rate myself, but that is the rate someone else
obtained from their experiments.  It's not my work and estimates on that so
I can't answer anything else about it.  Probably a liberal number.

			-- Aaron




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