ASAP PLEASE -- Cloning PCR fragments

berns001 at coyote.csusm.edu berns001 at coyote.csusm.edu
Sun Oct 22 18:30:59 EST 1995


98-bp fragments synthesized by PCR (using Vent).
Trying to clone into pTZ18R.
Vector cut with XmaI and treated with CIP.
PCR inserts cut at one end with XmaI; other end is whatever was PCR'd. 
Both vectors and inserts will be gel purified (0.7% Sea-Kem).
Ligation will take place in gel slices (only need one to work).

Should I phosphorylate the XmaI-cut PCR fragments prior to
gel purification?

Note:  The first ligation is only to connect the XmaI sites.  
I'll worry about circularization later.

Thanks, Debbie.



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