Adding restriction sites to PCR primers

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Tue May 16 08:53:14 EST 2000



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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