PCR: Why is my TAQ unfaithful
Julie Wilberding
jwilber at pop.nd.edu
Tue Nov 22 14:23:30 EST 1994
Hi,
I have been tying to PCR three constructs. I use Promega TAQ, 100uM
dNTP's, buffer and the proper amount of primers and template (Double
stranded circular plasmid). My mutation rate is high ( 3 in one 180 bp
fragment). I have changed dNTP, TAQ, with no change. This is a problem
recently encountered in our lab. The PCR conditions are 94 1 min., 55 1
min., 72 2 min., 30 cycles with a final cycle b3ing a 10 min extension at
72. Is the circular template a possible cause??? Efficiency is good but
fidelity sucks! Any help or advice would be greatly appreciated.
Julie
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