Ligation of PCR products

David Steffen steffen at mbir.bcm.tmc.edu
Fri Jul 19 10:01:24 EST 1991


(Michael Newstein) writes:
>I have read that it is exceedingly difficult to blunt-end ligate products from
> PCR  (without cleaving first in internal restriction sites).

Our experience is that it is exceedingly difficult to blunt-end ligate
PCR products without doing something to them.  That something can be
either cleaving with a restriction enzyme, as you suggest, or blunting
the ends with Klenow.  The method we use for Klenow blunting is
standard (probably straight from Maniatis).  If you need a detailed
method, email me.


-- 
David Steffen
Department of Cell Biology, Baylor College of Medicine, Houston TX 77030
Telephone = (713) 798-6655, FAX = (713) 790-0545
Internet = steffen at mbir.bcm.tmc.edu



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