digesting the ends of PCR products

[Steven Goldberg]

In article <32F24C5E.5A11 at sluvca.slu.edu>, tysondr at sluvca.slu.edu wrote:

> s j peake wrote:
> > 
> >    I've placed restriction sites in the ends of my PCR product by
engineering
> > the sites into my primers.  The BamH1 site at one end cuts okay, but
the Kpn1
> > site at the other end doesn't cut at all . Both sites have two base pairs to
> > act as a clamp which according to the NEB catalogue should be fine.  Has
> > anyone got any ideas or do l need to get a new primer?
> 

Another possible solution is to use a vector that can accept PCR fragments
(like pCRII) and get your fragment into a plasmid.  Then it should be easy
to digest and isolate the correct fragment for cloning into your plasmid
of choice, if needed.

Steve