Pwo polymerase blues

hassane mchaourab hassane at wlheye.jsei.edu
Thu Mar 27 21:37:15 EST 1997


Hi,

 I recently decided to try Pwo polymerase from Boehringer simply because
I got a discount and because the "propaganda litterature" indicated that
the enzyme has similar fidelity to pfu from stratagene. I used pwo for
site-directed mutagenesis by PCR. Initially, I noticed a significant
increase in the DNA yield  compared to pfu (great!!). However,  in both
clones I have sequenced, I found misincorporations. Now I admit that I
do not have a statistically significant set of data (only two clones),
but I am worried since in two years of using pfu, I have not found a
single instance of misincorporation. Please note that my amplified piece
is only 200 bp. Has anybody noticed or heard of a similar problem?



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