Simultaneous Genotyping of Single-nucleotide Polymorphisms in Alcoholism-related Genes Using Duplex and Triplex Allele-specific PCR with Two-step Thermal Cycles

被引：4

作者：

Shirasu, Naoto ^{[1
]}

Kuroki, Masahide ^{[1
]}

机构：

[1] Fukuoka Univ, Dept Biochem, Fac Med, Fukuoka 8140180, Japan

来源：

ANALYTICAL SCIENCES | 2014年 / 30卷 / 11期

关键词：

Alcohol dehydrogenase; aldehyde dehydrogenase; allele-specific PCR; genotyping; mu-opioid receptor; single nucleotide polymorphism; two-step PCR; JAPANESE; ASSOCIATION; DEPENDENCE; A118G; BLOOD;

D O I：

10.2116/analsci.30.1093

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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 mu-opioid receptor. Applying MightyAmp (R) 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.

引用

页码：1093 / 1096

页数：4

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