Simultaneous genotyping of single-nucleotide polymorphisms in alcoholism-related genes using duplex and triplex allele-specific PCR with two-step thermal cycles.

Abstract

We developed a time- and cost-effective multiplex allele-specific polymerase chain reaction (AS-PCR) method based on the two-step PCR thermal cycles for genotyping single-nucleotide polymorphisms in three alcoholism-related genes: alcohol dehydrogenase 1B, aldehyde dehydrogenase 2 and μ-opioid receptor. Applying MightyAmp(®) DNA polymerase with optimized AS-primers and PCR conditions enabled us to achieve effective and selective amplification of the target alleles from alkaline lysates of a human hair root, and simultaneously to determine the genotypes within less than 1.5 h using minimal lab equipment.

• Authors: Naoto Shirasu, Masahide Kuroki
• Journal: Analytical sciences : the international journal of the Japan Society for Analytical Chemistry (Anal Sci) Vol. 30 Issue 11 Pg. 1093-6 ( 2014) ISSN: 1348-2246 [Electronic] Switzerland
• PMID: 25382047 (Publication Type: Journal Article)

Chemical References

• Receptors, Opioid, mu
• Alcohol Dehydrogenase
• Aldehyde Dehydrogenase

Topics

• Alcohol Dehydrogenase (genetics)
• Alcoholism (genetics)
• Aldehyde Dehydrogenase (genetics)
• Alleles
• Base Sequence
• Cost-Benefit Analysis
• Genotyping Techniques (economics, methods)
• Polymerase Chain Reaction (economics, methods)
• Polymorphism, Single Nucleotide
• Receptors, Opioid, mu (genetics)
• Temperature
• Time Factors