Shen Gong-Qing, Abdullah Kalil G, Wang Qing Kenneth
Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA.
Methods Mol Biol. 2009;578:293-306. doi: 10.1007/978-1-60327-411-1_19.
Single nucleotide polymorphisms (SNPs) are common DNA sequence variations that occur at single bases within the genome. SNPs have been instrumental in elucidating the genetic basis of common, complex diseases using genome-wide association studies, candidate gene case-control association studies, and genome-wide linkage analyses. A key to these studies is genotyping of SNPs. Various methods for SNP genotyping have been developed. For a particular genotyping project, the choice of method is dependent on the number of SNPs (n) and the number of DNA samples (m) to be genotyped. For a genome-wide or large-scale project with very high n and small m, the Affymetrix SNP GeneChip and Illumina GoldenGate BeadChips assays are the ideal methods. For a project involving a small number of SNPs (small n) and a large population (high m), the TaqMan assay is the preferred technology as it has high throughput and is highly accurate, precise, time-efficient, and cost-effective. Here, we describe the detailed procedures for TaqMan SNP genotyping assay, including preparation of high-quality DNA samples, the operating protocol, clarification of technical issues, and discussion of several cautionary notes.
单核苷酸多态性（SNPs）是基因组内单个碱基处发生的常见DNA序列变异。利用全基因组关联研究、候选基因病例对照关联研究和全基因组连锁分析，SNPs在阐明常见复杂疾病的遗传基础方面发挥了重要作用。这些研究的关键是SNPs的基因分型。已经开发出了多种SNPs基因分型方法。对于特定的基因分型项目，方法的选择取决于要进行基因分型的SNPs数量（n）和DNA样本数量（m）。对于n非常高且m较小的全基因组或大规模项目，Affymetrix SNP基因芯片和Illumina GoldenGate微珠芯片检测是理想的方法。对于涉及少量SNPs（小n）和大量人群（高m）的项目，TaqMan检测是首选技术，因为它具有高通量，且高度准确、精确、省时且经济高效。在此，我们描述了TaqMan SNP基因分型检测的详细步骤，包括高质量DNA样本的制备、操作流程、技术问题的澄清以及几个注意事项的讨论。
