微测序技术检测12个Y-SNP及其遗传多态性

A Minisequencing Technique for Genotyping 12 Y-SNP and it′s Genetic Polymorphisms

目的应用SNaPshotKit对Y染色体上12个SNP位点进行快速而准确的检测,对四川地区78个汉族男性无关个体进行群体遗传学研究,并对陈旧骨骼和性犯罪案件的相关物证进行检验。方法对SRY2627、SRY1532、M13、M20、SRY8299、Tat、M69及M9、92R7、M17、M19、M112两组共12个Y-SNP位点进行复合扩增,PCR产物经纯化处理后,采用SNaPshotKit试剂结合毛细管电泳技术对单核苷酸多态性进行检测。结果建立了12个Y-SNP位点的微测序快速检测系统,在四川地区人群中发现M9、SRY8299二个位点存在变异。结论复合扩增结合微测序技术能够同时对多个Y-SNP的多态性进行快速而准确的检测,建立的检测系统在法医学个体识别中具有应用价值。

Objective The aim of the present study was to establish a rapid and robust assay used to simultaneously genotype SNPs by the single nucleotide primer extension （minisequencing） with the SNaPshot Kit and obtain the population genetic data in Chinese population in Sichuan. The analysis of single nucleotide polymorphisms （SNPs） is a promising application in forensic casework. Methods 12 Y-SNPs, which were SRY2627, SRY1532, M13, M20, SRY8299, Tat, M69, M9, 92R7, M17, M19 and M112, were multiple amplificated and the PCR products were pooled , Purified, and then used as templates for the minisequencing reaction with the commol/Lercially available SNaPshot Kit. Then the products of minisequencing reaction were detected by capillary, elcetrophoresis. Results 78 genomic DNA individual samples from Sichuan, China and 5 semen stain samples from sexal criminal scene were analyzed and two haplotypes could be identified. Conclusion A rapid method has been established to analyze the 12 Y-SNPs by multiplex PCR and minisequencing. It can be applied in forensic casework successfully.