TA cloning of a PCR product generate by Vent DNA polymerase

Dr. Yu zyu at unixg.ubc.ca
Tue Jul 29 22:52:30 EST 1997


Does anyone has any successful experience in cloning a PCR product
generated by Vent DNA polymerase? The product is 2.7 kbp.  I followed
the 
InVitrogen protocol and incubate the Vent PCR product in Taq at 72 degC
for 10 minutes, phenol extract and preciptate the DNA at room temp. The
precipitated DNA is resuspended in TE and immediate ligate to the TA
vector. I tried this several times but I don't get any clones.

Tai Man Louie
Dept. of Microbiology,
Univ. of British Columbia



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