Targeted SNP Genotyping Using the TaqMan® Assay
Targeted SNP Genotyping Using the TaqMan® Assay
复制标题
DOI：
10.1007/978-1-61737-954-3_6
复制
发表时间：
2011-01-01
期刊：
影响因子：
--
通讯作者：
Kovacs, Peter
中科院分区：
文献类型：
--
作者：
Schleinitz, Dorit;DiStefano, Johanna K.;Kovacs, Peter
超过99%的基因组DNA序列在人类中是相同的，并且毫不奇怪，序列中的微小变化通常会对表型产生重大影响。序列变体还可以介导人类对疾病易感或对环境因素如细菌、病毒、毒素、化学品、药物和治疗干预作出反应的方式。单核苷酸多态性（SNP）是当基因组序列中的单个碱基可以由至少两个不同的核苷酸表示时发生的DNA序列变异。在过去的十年中，已经确定了许多SNP，这些SNP至少部分地解释了复杂疾病的遗传结构，如癌症，糖尿病，血管并发症，某些形式的精神疾病和许多其他疾病。疾病相关的SNP通常通过候选基因方法或最近通过全基因组关联研究来鉴定。在任何一种情况下，相关性的发现都需要在独立的、基于人群的研究样本中进行验证，这些样本通常由数百/数千人组成。在这类研究中对中等数量的标记进行基因分型的便利技术可利用TaqMan平台（Applied Biosystems; Foster City，CA）获得，其利用聚合酶链反应扩增和等位基因辨别以成本有效的方式容易且有效地产生基因型数据。在这里，我们介绍和描述这种常用的技术，并包括协议，可以直接用于实验室，旨在进行中到大规模的基因分型研究。
More than 99% of genomic DNA sequence is identical among humans, and not surprisingly, slight variations in sequence can often produce a major effect on phenotype. Sequence variants may also mediate the manner in which humans are susceptible to disease or respond to environmental factors such as bacteria, viruses, toxins, chemicals, drugs, and therapeutic interventions. Single-nucleotide polymorphisms (SNPs) are DNA sequence variations that occur when a single base in the genome sequence can be represented by at least two different nucleotides. In the last decade, numerous SNPs have been identified that explain, at least partially, the genetic architecture of complex diseases such as cancer, diabetes, vascular complications, some forms of mental illness, and a multitude of other disorders. Disease-related SNPs are commonly identified through candidate gene approaches, or more recently, through genome-wide association studies. In either case, findings of association require verification in independent, population-based, study samples, usually consisting of several hundreds/thousands of individuals. A convenient technique to genotype a moderate number of markers in this kind of study is available with the TaqMan (R) platform (Applied Biosystems; Foster City, CA), which utilizes polymerase chain reaction amplification and allelic discrimination to easily and efficiently generate genotype data in a cost-effective way. Here, we introduce and describe this commonly used technique and include protocols that can be directly used in laboratories aiming to perform moderate- to large-scale genotyping studies.