Enhanced single-strand conformation polymorphism (SSCP) detection of point mutations utilizing methylmercury hydroxide

Weghorst, C.M.; Buzard, G.S.

Biotechniques 15(3): 396-8 400


ISSN/ISBN: 0736-6205
PMID: 8217149
Accession: 002370175

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Single-strand conformation polymorphism (SSCP) analysis of PCR amplified products to detect single-base substitutions involves denaturating the PCR fragments, followed by polyacrylamide gel electrophoresis under non-denaturing conditions. This paper reports the use of methylmercury hydroxide as a strong denaturing agent to enhance strand separation. The technique was evaluated on point mutations in rat mesenchymal tumours. The results demonstrated that the addition of methylmercury hydroxide to the SSCP protocol reduced the complexity of electrophoretic banding patterns by eliminating artifacts resulting from partial DNA fragment denaturation.