Genotyping of alcohol dehydrogenase type 2 and 3 using a two-buffer polyacrylamide gel electrophoresis system

Clin Chem Lab Med. 2003 Mar;41(3):298-301. doi: 10.1515/CCLM.2003.047.

Abstract

Genetic polymorphisms in the alcohol dehydrogenase genes, ADH2 and ADH3, have been shown to affect individual susceptibility to diseases such as alcoholism and oesophageal cancer. Although several PCR-based methods for genotyping these enzymes have been previously developed, the two-buffer polyacrylamide gel electrophoresis system has the ability to rapidly resolve all classes of point mutations within 2-3 hours instead of the conventional overnight separation. The success of this technique is partly attributable to a discontinuous two-phase buffer system and horizontal flatbed gel electrophoresis rather than conventional vertical gels. Using a modification of this system, we were able to detect all of the known polymorphisms within ADH2 exons 3 and 9 and ADH3 exon 8, as well as a rare variant within ADH2 exon 9. This method is rapid, cost-effective, and is ideal for large scale screening programmes.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Dehydrogenase / genetics*
  • Aldehyde Oxidoreductases / genetics*
  • Buffers
  • DNA / blood
  • DNA Primers / chemistry
  • Electrophoresis, Polyacrylamide Gel / methods
  • Exons / genetics
  • Genotype
  • Humans
  • Polymerase Chain Reaction
  • Polymorphism, Genetic*
  • Polymorphism, Single-Stranded Conformational
  • South Africa

Substances

  • Buffers
  • DNA Primers
  • DNA
  • Alcohol Dehydrogenase
  • formaldehyde dehydrogenase (glutathione)
  • Aldehyde Oxidoreductases