restriction digestion of PCR product for cloning

Paul Rohde P.Rohde at mailbox.uq.edu.au
Sat Oct 4 04:59:28 EST 1997


s9401501 at COUGAR.VUT.EDU.AU (Matthew Knight) writes: > I am having problems digesting a PCR product which has restriction enzyme
> sites placed into the PCR primers, thus they are very close to the end of
> the PCR product.
> 
> Does anyone have any suggestions on how I can be sure that the
> restriction enzyme is going to cut the PCR product. And will the
> restriction enzyme cut with the sequence being so close to the end of the
> product (i.e. about 5 or 6 bases)
> 
> Thanks in advance
> 
> Matthew Knight

As a guide check out New England Biolabs table on p. 238 96/97 issue,
 or look in index for "cleavage of short oligoes" under "Restriction
endonucleases"

Hope this will help.



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