Woodward John
DuPont Pioneer, 1000, 7100 NW 62nd Ave, Johnston, IA, 50131-0184, USA,
Methods Mol Biol. 2014;1145:67-74. doi: 10.1007/978-1-4939-0446-4_6.
With TaqMan(®) technology allele-specific probes are utilized for quick and reliable genotyping of known polymorphic sites. TaqMan assays are robust in genotyping multiple variant types, including single nucleotide polymorphisms, insertions/deletions, and presence/absence variants. To query a single bi-allelic polymorphism, two TaqMan probes labeled with distinct fluorophores are designed such that they hybridize to different alleles during PCR-based amplification of a surrounding target region. During the primer extension phase of PCR, the 5'-3' exonuclease activity of Taq polymerase cleaves and releases the fluorophores from bound probes. At the end of PCR, the emission intensity of each fluorophore is measured and allele determination at the queried site can be made.
利用TaqMan（®）技术，等位基因特异性探针可用于已知多态性位点的快速、可靠基因分型。TaqMan分析在对多种变异类型进行基因分型时表现稳健，包括单核苷酸多态性、插入/缺失以及存在/缺失变异。为检测单个双等位基因多态性，设计了两种用不同荧光团标记的TaqMan探针，使其在基于PCR的周围靶区域扩增过程中与不同等位基因杂交。在PCR的引物延伸阶段，Taq聚合酶的5'-3'核酸外切酶活性会切割并从结合的探针上释放荧光团。在PCR结束时，测量每个荧光团的发射强度，从而确定所检测位点的等位基因。
