PCR cloning

Miguel Valvano mvalvano at mni.uwo.ca
Thu Jul 4 11:12:28 EST 1996

Nitin S. Baliga wrote:
> It's becoming a rather well known fact that PCR products are difficult to
> clone. I wonder if anyone has found out ways to get around this problem,
> since PCR is a very powerful technique, and use of this technique may get
> limited to analytical research if cloning continues to be a problem!
> Nitin

We are quite successful in cloning PCR fragments by using PwoI 
polymerase. The basic protocol involves amplification, kinase treatment 
(if vector treated with alkaline phosphatase, and then ligation.  

Miguel A. Valvano, M.D., Associate Professor,
Department of Microbiology and Immunology
University of Western Ontario
London, Ontario, N6A 5C1, Canada
Phone (519) 661-3996/3433
Fax (519) 661-3499

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