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

Biotechniques. 2000 Sep;29(3):500-4, 505-6. doi: 10.2144/00293st01.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Technical Report

MeSH terms

  • Electrophoresis / methods*
  • Genotype*
  • Humans
  • Image Processing, Computer-Assisted
  • Insulin / genetics
  • Mutagenesis
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Polymorphism, Single Nucleotide / genetics*
  • Promoter Regions, Genetic
  • Software
  • alpha 1-Antitrypsin / genetics

Substances

  • Insulin
  • alpha 1-Antitrypsin