Yang Heyi, Wang Haijian, Wang Jie, Cai Yun, Zhou Gangqiao, He FuChu, Qian Xiaohong
Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, Beijing, 100850, PR China.
Anal Biochem. 2003 Mar 1;314(1):54-62. doi: 10.1016/s0003-2697(02)00641-3.
A robust high-throughput single-nucleotide polymorphism (SNP) genotyping method is reported, which applies allele-specific extension to achieve allelic discrimination and uses matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to measure the natural molecular weight difference of oligonucleotides for determination of the base in a single-nucleotide polymorphic location. Tenfold PCR is performed successfully by carefully designing the primers and adjusting the conditions of PCR. In addition, two ways used for PCR product purification are compared and the matrix used in mass spectrometry for high-throughput oligonucleotide analysis is evaluated. The result here shows that the method is very effective and suitable for high-throughput genotyping of SNPs.
报道了一种稳健的高通量单核苷酸多态性（SNP）基因分型方法，该方法应用等位基因特异性延伸实现等位基因鉴别，并使用基质辅助激光解吸/电离飞行时间质谱来测量寡核苷酸的天然分子量差异，以确定单核苷酸多态性位点的碱基。通过精心设计引物和调整PCR条件，成功进行了10倍PCR。此外，比较了两种用于PCR产物纯化的方法，并评估了用于高通量寡核苷酸分析的质谱基质。此处结果表明该方法非常有效，适用于SNP的高通量基因分型。
