TA cloning problems!!!!

Robert Fisher fisher at radonc.unc.edu
Fri Jul 21 11:29:37 EST 1995


Has anyone had problems TA cloning PCR products into a SmaI digested plasmid?

Briefly, I digested Bluescript II SK + with SmaI, T-tailed it with Taq
polymerase,  and ligated my genecleaned pcr product in with T4 DNA ligase.

Alas, no colonies.  I have been told that this is a good protocol, and I was
wondering if the problem was due to the digest, since I have heard  iffy things
about ligations into SmaI sites.




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