Cloning PCR products

Robert J. Coelen robert at
Thu Jun 4 22:23:33 EST 1992

In article <1992Jun3.160305.12666 at> naffara at (Dr. N.A. Affara) writes:
>This is a repeat of an earlier message which I think didn't get through,
>I apologise if you have seen this before.
>Has anyone tried to cut a DNA fragment (not a PCR product) with an enzyme
>which is near to the end e.g. up to 25 bases away. I am interested to
>find out if the problem cutting PCR products is because the site is too
>close to the end or something else entirely.

 25 nt from the end is in my book not really near the end. If you are within 
a couple of bases or so things might get a bit hairy, depending on the 
enzyme you use, however 25 nts should be no problem. We regularly open 
cloning vectors with a couple of enzymes, the second cut is often within the 
25 nt you mention.
Robert J. Coelen                               | phone : 09 - 389 3915
Department of Microbiology                     | fax   : 09 - 389 2912
The University of Western Australia            | 

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