Problems cutting PCR product

Duncan Clark junk at hgmp.mrc.ac.uk
Tue Sep 10 11:07:46 EST 2002


Historians believe that in newspost 
<3D7DA8C4.9080505 at chembio.REMOVETHISBEFOREREPLYING.ntnu.no> on Tue, 10 
Sep 2002, Trond Erik Vee Aune 
<trondaun at chembio.REMOVETHISBEFOREREPLYING.ntnu.no> penned the following 
literary masterpiece:
>> Two possibilities I suppose,
>>  1. Most likely, your restriction site is at the extreme end of your
>> PCR fragment and can therefor not cut. Remedy. Design primers with an
>> extra tail of irelevant nucleiotides or, easier in this case,
>> posphrylate/ligate your PCR fragment to make your site internal and
>> than cut , this should always work.
>
>
>No, the sites are designed so to be 25 nts from the ends of the fragment.

LIgate PCR product to itself and see if ligated product cuts with your 
RE's.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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