title: Multiplex SSCP and heteroduplex analysis with Southern hybridization for large-scale mutation detection.
abstract: We have developed a modification of the single-strand conformational analysis and heteroduplex analysis methods of mutation detection, with the intention of applying them to genetic diseases involving large genes or multiple genes producing a similar phenotype. The technique involves electrophoresing up to 10 or more DNA fragments on a polyacrylamide gel, followed by bidirectional Southern blotting and individual examination by hybridization. This can reduce the time involved in mutation detection by more than 50%. We confirmed the validity of our approach by detecting 90% of mutations in a blind study of previously characterized mutations in the adenomatous polyposis coli (APC) gene that underlies familial adenomatous polyposis.
authors.affiliation: Department of Human Molecular Genetics, University of Newcastle-upon-Tyne, Ridley Building, Claremont Road, Newcastle-upon-Tyne, NE1 7RU, United Kingdom. robert.pogue@ncl.ac.uk
authors.forename: R
authors.initials: R
authors.lastname: Pogue
authors.forename: S
authors.initials: S
authors.lastname: West
authors.forename: K
authors.initials: K
authors.lastname: Bushby
mesh_terms: D011125:Adenomatous Polyposis Coli / Q000235:genetics*; D015139:Blotting, Southern; D004252:DNA Mutational Analysis / Q000379:methods*; D017491:Genes, APC / Q000235:genetics*; D020180:Heteroduplex Analysis; D006801:Humans; D018807:Polymorphism, Single-Stranded Conformational