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目的建立常染色体21个SNPs的多态性分型方法。方法采用荧光标记公用引物和等位基因特异性引物原理设计SNP复合扩增引物体系,对45个备选SNP位点筛选,选出21个及性别Amelogenin构成复合扩增体系。PCR产物经3130XL型电泳仪电泳分离,GeneMaperTM3.0数据分析软件分析结果。同时随机选取6份样品,使用测序方法对SNP分型并进行测序验证。结果应用本研究建立的复合扩增体系扩增样品,产物经毛细管电泳后,每个SNPs均可正确判定基因型。随机选取6份样品SNPs位点测序结果显示,荧光标记SNPs复合扩增分型与直接测序结果完全一致。结论本研究建立的荧光标记公有引物特异性片段常染色体21个SNPs复合扩增方法是SNP多态性分析的一种有效方法,并有助于解决SNP分型识别能力、效率、通量和高成本的问题。
Objective To establish a polymorphism typing method of 21 SNPs in autosomes. Methods SNP amplification primers were designed by using fluorescent primers and allele-specific primers. A total of 45 candidate SNPs were screened, and 21 and Amelogenin were selected to form a composite amplification system. PCR products were electrophoresed on a 3130XL electrophoresis system, and the results were analyzed with GeneMaperTM3.0 data analysis software. At the same time, six samples were randomly selected and SNP was typed and sequenced using sequencing. Results Using the multiplex amplification system established in this study to amplify samples, the genotype of each SNP can be correctly determined by capillary electrophoresis. Sequence analysis of SNPs in randomly selected six samples showed that the SNP genotyping of fluorescently labeled SNPs was completely consistent with the direct sequencing results. Conclusion The 21 common SNPs amplified by the fluorescent-labeled common primer-specific fragments in this study could be an effective method for SNP polymorphism analysis and help to resolve the SNP typing ability, efficiency, flux and high The cost of the problem.