title: Duplex PCR-RFLP for the Simultaneous Genotyping of Single Nucleotide Polymorphisms in ADH1B and ALDH2 Genes.
abstract: Single nucleotide polymorphisms (SNPs) in ADH1B and ALDH2, which encode alcohol dehydrogenase 1B and aldehyde dehydrogenase 2, respectively, are responsible for ethanol-metabolizing activity and alcohol-related disease. We developed a simple and rapid polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method for the simultaneous genotyping of the genes in a single-tube reaction. Using an alkaline lysate of a human hair root as a template, SNP-containing regions of each gene were PCR-amplified simultaneously. The PCR products were directly applied to double restriction digestion, followed by agarose gel band analysis. The duplex PCR-RFLP allows for the simultaneous and accurate determination of the SNP genotypes of ADH1B and ALDH2 within 1.2 h.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Fukuoka University.
authors.forename: Naoto
authors.initials: N
authors.lastname: Shirasu
authors.forename: Shin'ichiro
authors.initials: S
authors.lastname: Yasunaga
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000071396:Aldehyde Dehydrogenase, Mitochondrial / Q000235:genetics*; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D013997:Time Factors
chemical_list: C489332:ADH1B protein, human; D000426:Alcohol Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial