Adding restriction sites to PCR primers
l_shen9 at yahoo.com
Tue May 16 11:56:40 EST 2000
--- Susanne Rohrer
> Beverly Metcalfe wrote:
> > I am designing primers and I want to add
> restriction enzyme sites to the
> > 5'ends. To do this, do I just add the restriction
> site to the 5' end, or do
> > I need to add extra bases after the enzyme's site
> so the the enzyme will be
> > able to cleave it?
> depends on the enzyme - look it up in the back of
> the New England Biolabs
> Catalogue, or get the same information on their
> website (www.neb.com). Some
> will not cut very well. For digestion of PCR ends,
> I recomment you do it
> overnight in an incubator.
> Susanne Rohrer
> Institute of Medical Microbiology
> University of Zurich
> srohrer at immv dot unizh dot ch
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