使用TaqMan PCR对三等位基因单核苷酸多态性进行基因分型。

Genotyping of triallelic SNPs using TaqMan PCR.

作者信息

Morita Akihiko, Nakayama Tomohiro, Doba Nobutaka, Hinohara Shigeaki, Mizutani Tomohiko, Soma Masayoshi

机构信息

Department of Medicine, Divisions of Neurology, Nihon University School of Medicine, Tokyo, Japan.

出版信息

Mol Cell Probes. 2007 Jun;21(3):171-6. doi: 10.1016/j.mcp.2006.10.005. Epub 2006 Nov 7.

DOI:10.1016/j.mcp.2006.10.005

PMID:17161935

Abstract

Methods for analysis of single nucleotide polymorphisms (SNP) are well developed. However, most ready-made SNP genotyping kits (e.g., those that use TaqMan PCR) are based on the assumption that the SNP is biallelic. Thus, most kits are unsuitable for triallelic SNPs. We have experienced difficulty genotyping an SNP using TaqMan PCR. In the present study, we developed a method of genotyping a triallelic SNP (rs3091244: alleles C, A and T) using TaqMan PCR. We used 2 different genotyping kits: one for C/A allele genotyping, and one for C/T allele genotyping. The results of these 2 kits were combined to complete the genotyping. The subjects were 320 essentially healthy elderly Japanese. The frequencies of the C, A and T alleles were 0.78, 0.155 and 0.065, respectively. This double-tube method using TaqMan SNP Genotyping Assays was very accurate and convenient, and it should be useful for genotyping in case-control association studies or linkage studies.

摘要

单核苷酸多态性（SNP）的分析方法已经很成熟。然而，大多数现成的SNP基因分型试剂盒（例如那些使用TaqMan PCR的试剂盒）都是基于SNP是双等位基因的假设。因此，大多数试剂盒不适用于三等位基因SNP。我们在用TaqMan PCR对一个SNP进行基因分型时遇到了困难。在本研究中，我们开发了一种使用TaqMan PCR对三等位基因SNP（rs3091244：等位基因C、A和T）进行基因分型的方法。我们使用了2种不同的基因分型试剂盒：一种用于C/A等位基因分型，另一种用于C/T等位基因分型。将这2种试剂盒的结果结合起来以完成基因分型。研究对象为320名基本健康的日本老年人。C、A和T等位基因的频率分别为0.78、0.155和0.065。这种使用TaqMan SNP基因分型检测的双管法非常准确且方便，它应该对病例对照关联研究或连锁研究中的基因分型有用。