Separating cut from uncut vector

Steven Enkemann enkemans at dc37a.nci.nih.gov
Mon Sep 4 15:53:49 EST 1995


In article <jpcd0-0409951419190001 at macr1-1.welc.cam.ac.uk>,
jpcd0 at mole.bio.cam.ac.uk (John Dixon) asked:

> what's a Hapaxoterministic Enzyme?

in response to my posting:
>  
> > 
> > Hey guys why bother trying to isolate the uncut vector from the cut
> > vector.  Instead chop up the vector further with a Hapaxoterministic
> > Enzyme.  They are readily available and when used can make these kids of
> > cloning problems non-existant.  For references on how to use them see:
> > Berger et al. (1993) Analytical Biochemistry 214: 571-579. and Berger,
> > S.L. (1994) Analytical Biochemistry 222: 1-8.
> > 
> 
> 
They are restriction enzymes that cut outside of their recognition
sequence so that the termini they produce are unique for each cut site. 
An example is Sfi I that cuts ggccnnnn/nggcc.  A plasmid can be cut into
several pieces by one of these enzymes and yet it can be ligated back
together in its original form because the ends only go back together one
way.  For more information check the references I cited.

Steven A. Enkemann PhD.

I don't believe that curiosity killed the cat.  
I think it just did the wrong experiment.



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