- Step by Step SSCP (Travis Glenn, Laboratory of Molecular Systematics, Smithsonian Institution)
This protocol is for Single-Strand Conformation Polymorphism (SSCP) analysis. It is basically the same as any SSCP protocol you will find in the literature. The only major difference is that the author recommends doing a two reaction PCR (unless you are screening more than 48 individuals, then you should optimize PCR conditions and amplify hot the first time around). The first reaction is a "normal" cold PCR to amplify the intended products. Check that these reactions were successful by running out on a minigel. The second reaction is a hot PCR to label the products produced in the first PCR reaction. The hot product will be diluted in sequencing buffer, denatured, and loaded onto a native polyacrylamide gel.
Added: Mon Sep 23 2002, Hits: 978, Reviews: 0
Mutation Detection By Single-Strand Conformational Polymorphism (SSCP)
(Reddy research laboratory, Neurological Sciences Institute, Oregon Health & Science University)
PCR Method for mutation detection.
Added: Mon Sep 23 2002, Hits: 930, Reviews: 0