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Biotechniques. 2000 Sep;29(3):500-4, 505-6.

SNP genotyping by combination of 192-well MADGE, ARMS and computerized gel image analysis.

Author information

1
Human Genetics Research Division, Southampton University Medical School, Southampton General Hospital, England, UK. s.d.o'dell@soton.ac.uk

Abstract

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.

PMID:
10997263
DOI:
10.2144/00293st01
[Indexed for MEDLINE]

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