See D, Kanazin V, Talbert H, Blake T
Montana State University, Bozeman, USA.
Biotechniques. 2000 Apr;28(4):710-4, 716. doi: 10.2144/00284st07.
Single-nucleotide polymorphisms (SNPs) represent the most prevalent class of genetic markers available for linkage disequilibrium or cladistic analyses. PCR primers may be labeled with fluorescent dyes and used to rapidly and accurately differentiate among alleles that are defined by a single-nucleotide differences. Here, we describe the primer-mediated detection of SNPs based on primer mismatch during allele-specific amplification of preamplified target sequences. Primers are labeled with different fluors at their 5' nucleotides, with their 3' termini at the transition mutation that defines allelic variation at the target locus. Each primer perfectly matches one of the two available alleles for each locus. Electrophoretic detection permits characterization of the product both by size and fluor. This report demonstrates some of the capabilities of this assay, including heterozygote determination and multiplexed analysis.
单核苷酸多态性（SNPs）是用于连锁不平衡或系统发育分析的最普遍的一类遗传标记。聚合酶链反应（PCR）引物可用荧光染料标记，并用于快速、准确地区分由单核苷酸差异定义的等位基因。在此，我们描述了基于预扩增靶序列等位基因特异性扩增过程中引物错配的单核苷酸多态性引物介导检测方法。引物在其5'核苷酸处用不同的荧光标记，其3'末端位于定义靶位点等位基因变异的转换突变处。每个引物与每个位点的两个可用等位基因之一完美匹配。电泳检测允许通过大小和荧光对产物进行表征。本报告展示了该检测方法的一些能力，包括杂合子测定和多重分析。
