Multiplex SSCP and heteroduplex analysis with southern hybridization for large-scale mutation detection

• Autor(a) principal: Pogue, Robert Edward
• Data de Publicação: 1998
• Outros Autores: West, S., Bushby, Katharine M. D.
• Tipo de documento: Artigo
• Idioma: por
• Título da fonte: Repositório Institucional da UCB
• Texto Completo: http://twingo.ucb.br:8080/jspui/handle/10869/637; https://repositorio.ucb.br:9443/jspui/handle/123456789/7804

We have developed a modification of the singlestrand 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.