Problems cutting PCR product
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
>> 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
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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