blunt-end ligation with PCR product?

Paul Dunman dunman at njmsa.UMDNJ.EDU
Fri Jun 20 10:53:29 EST 1997


Minh Luu <pluu at students.wisc.edu> writes:

>Help,

>I'm trying to clone some PCR products, from pfu polymerase which
>generates blunt-ended products, into a plasmid cut with Ecl136II.  I've
>tried phosphatase treatment, adding PEG into my ligation, all without
>any success because all I see in my minipreps are false-positives
>(deletions mostly).  I'm thinking about getting Statagene's pCR-Script
>kit.  Can anyone offer some insight?  Thanks

>Minh

You didn't mention...did you phosporylate your PCR products prior to 
attempting ligation?

Just curious.

Paul D.



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