TA cloning nightmare

Margon Vandongen vando006 at mc.duke.edu
Tue Mar 12 11:06:48 EST 1996


Catherine,

For what it is worth: we routinely use PFU (and sometimes a mixture of
Taq and PFU) to amplify our fragments. Our experience is that you get a
very low efficiency in TA cloning unless you add an extra step to 
"taq-on" A's. We spin the product through a column, add Taq buffer and 
dNTP's and Taq polymerase and incubate at 72 for 10 minutes.It is a 
little elaborate but worth the trouble works like a charm!! Good Luck,

Margon





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