SNP genotyping by combination of 192-well MADGE, ARMS and computerized gel image analysis.
A new modification of the microplate array diagonal gel electrophoresis (MADGE) system accommodates the dual amplification refractory mutation system (ARMS) 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. Customized software by Phoretix 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/HphI 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.
O'Dell
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Human Genetics Research Division, Southampton University Medical School, Southampton General Hospital, England, UK. s.d.o'dell@soton.ac.uk
Gaunt
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D004586:Electrophoresis / Q000379:methods*; D005838:Genotype; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007328:Insulin / Q000235:genetics; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012984:Software; D000515:alpha 1-Antitrypsin / Q000235:genetics
D007328:Insulin; D000515:alpha 1-Antitrypsin