title: Genotyping of alcohol dehydrogenase type 2 and 3 using a two-buffer polyacrylamide gel electrophoresis system.
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.
authors.affiliation: MRC/UCT Oesophageal Cancer Research Group, Institute for Infectious Diseases and Molecular Medicine, Division of Medical Biochemistry, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa. rballo@curie.uct.ac.za
authors.forename: Robea
authors.initials: R
authors.lastname: Ballo
authors.forename: Dong-Ping
authors.initials: DP
authors.lastname: Li
authors.forename: M Iqbal
authors.initials: MI
authors.lastname: Parker
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000445:Aldehyde Oxidoreductases / Q000235:genetics*; D002021:Buffers; D004247:DNA / Q000097:blood; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005091:Exons / Q000235:genetics; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D013019:South Africa
chemical_list: D002021:Buffers; D017931:DNA Primers; D004247:DNA; D000426:Alcohol Dehydrogenase; C020145:formaldehyde dehydrogenase (glutathione); D000445:Aldehyde Oxidoreductases