SSCP analysis

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Fri May 28 12:17:12 EST 1993


In article <9305251750.AA19694 at net.bio.net>, JOFTOPSM at VTVM1.CC.VT.EDU (Donna Jensen) writes:
> Hi!
> Can anyone out there give me any information on Single-Stranded
> Conformation Polymorphism (SSCP) analysis for detecting point
> mutations?  I am interested in possibly using this technique to
> detect point mutations in bacteria.  I would appreciate any information,
> including references on the methods and principles involved in this
> technique, as well as information from those of you that have used SSCP
> on how well the technique works and what its limitations are.
> Thanks in advance!! :->
> Donna
> JOFTOPSM at VTVM1.CC.VT.EDU

I have used SSCP analysis before.  I had identified some point mutations in
human topo II alpha by PCR and sequencing, and I used the "known" mutant PCR
products to get the SSCP system working.  (I also used them for chemical
mismatch cleavage, but that is another story.)  The idea was promising for
detection of mutants in patient samples, but I had other fish to fry, so Dr.
Mary Danks in our department took over the project.  We have published a paper
in Cancer Research 53:1373-1379 (1993) "Single-strand conformational
polymorphism analysis of the Mr 170,000 isozyme of DNA topoisomerase II in
human tumor cells".  This paper gives all of the references which would be of
the most use to someone just starting out.  The primary reference is Orita, M.;
Iwahana, H.; Kanazawa, H.; Hayaski, K. and Kekiya, T. "Detection of
polymorphisms of human DNA by gel electrophoresis and single-strand
conformation polymorphisms"  PNAS 86:2766-2770 (1989).  If you would like a
reprint of the Danks, et al. paper, please let me know.  
SSCP is an excellent technique and is not too difficult to get going.  It is
conducive to "screen"-type labs, where someone sets up the assay and anyone
following the "recipe" can do it.  Good luck!
Barbara Bugg
Molecular Pharmacology
St. Jude Children's Research Hospital
Memphis, TN USA  38105



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