Simultaneous genotyping of alcohol dehydrogenase 2 and aldehyde dehydrogenase 2 by single‐strand conformation polymorphism analysis

Abstract

Alcohol dehydrogenase (ADH; EC 1.1.1.1) and aldehyde dehydrogenase (ALDH; EC 1.2.1.3) have important roles in the elimination of ingested ethanol. These enzymes have polymorphisms resulting from single‐point mutations that cause kinetic differences in their respective enzyme activities. Simultaneous observation of these enzymes would be useful in investigating the association between these enzyme polymorphisms and alcohol‐related problems. In this study amplified genomic DNA was amplified from nail clippings with two sets of primers for ADH2 and ALDH2 genes, respectively, in a micro test tube and the accuracy of the amplification was verified by direct sequencing. The PCR products were separated into four distinct bands by single‐strand conformation polymorphism analysis. This genotyping method is fast, accurate, reliable and inexpensive, and requires the same amount of template DNA as non‐simultaneous methods. In other words, the required amount of template DNA for this method is only half that required for the separate genotyping of ADH2 and ALDH2.

• Direct sequencing
• electrophoresis
• genomic DNA from nails
• PCR