Finding restrictase by recognition sequence

Peter Wang plw at mrc-lmb.cam.ac.uk
Tue Feb 23 06:16:13 EST 1999


Max Myakishev wrote:

> I know a point substitution (SNP) in a certain gene.
> For example,
> acgtgt(G/A)aatatcga
> I would like to design a PCR/RFLP assay for genotyping this SNP.

Dear Max:

A few restriction polymorphism ideas for the sequence you mentioned.  (I
hope it's really the one you're interested in?)  I couldn't find an enzyme
that distinguishes this without some additional mutation, but mismatching
primers can introduce them.  This means the size difference between cut and
uncut will mainly depend on the length of the primer used.

   AC GTGT(G)AATATCGA
CAYNN/NNRT G = recognition site for MslI

 AC GT GT(G)AATATCGA
CACNNN/GT G = recognition site for DraIII

ACGTGT(G)AATATCGA
       G AAT TC = recognition site for EcoRI

ACGTGT(G)AATA TCGA
       G AANN/NNTTC = recognition site for XmnI

Cheers,
- Peter

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Peter Wang, M.D., Ph.D.
MRC Centre for Protein Engineering,
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