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
> > the sites into my primers. The BamH1 site at one end cuts okay, but
> > 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.