Adding restriction sites to PCR primers

Susanne Rohrer "srohrer(removethis)" at
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 (  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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