SNP Genotyping by Combination of 192-Well MADGE, ARMS and Computerized Gel Image Analysis. (September 2000)

• Record Type: Journal Article
• Authors: O'Dell, Sandra D.; Gaunt, Tom R.; Day, Ian N.M.
• Is Part Of: Biotechniques. Volume 29:Number 3(2000)
• Page Start: 500
• Page End: 506
• Publication Date: 2000-09
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/00293st01
• Languages: English
• ISSNs: 0736-6205

Abstract

A new modification of the microplate array diagonal gel electrophoresis (MADGE) system accommodates the dual amplification refractory mutation system (ARMS TM ) products of 96 samples on one 192-well gel. Simultaneous electrophoresis of a number of horizontal ARMS-MADGE gels achieves high throughput. Gels are imaged digitally, here using the FluorImager ® 595 fluorescent scanning system. Customised software by Phoretix TM enables rapid computerized calling of band patterns in ARMS-MADGE arrays, in which the two wells receiving a pair of allele-specific assays for a single template are juxtaposed to form one virtual track, with genotype data exported directly into Microsoft ® Excel ® for statistical analysis. An ARMS assay of the A/T base change at the −23/ Hph I RFLP in the insulin gene promoter, which initiates from 2.5 ng template DNA, was used here to demonstrate this improved general approach for population SNP analyses.