Adding restriction sites to PCR primers

LI SHEN l_shen9 at yahoo.com
Tue May 16 11:56:40 EST 2000


--- Susanne Rohrer
<"srohrer(removethis)"@immv.unizh.ch> wrote:
> 
> 
> 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
> Switzerland
> 
> srohrer at immv dot unizh dot ch
> 
> 


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