GITRL基因SNP-rs2236876分型方法的比较

Comparison of method for GITRL SNP-rs2236876 genotyping

目的:对3种单核苷酸多态(single nucleotide polymorphism,SNP)分型方法进行比较,选择准确有效的SNP分型方法对GITRL SNP-rs2236876A/G进行基因分型。方法:采用Taqman-MGB探针法,限制性片段长度多态性聚合酶链式反应(cleaved amplification polymorphism sequence-tagged sites,PCR-RFLP)法和直接测序法对GITRL SNPrs2236876A/G进行分型并比较。结果:PCR-RLFP法分型结果直接通过电泳图可见,适合少量的实验对象;TaqmanMGB探针法通过不同的荧光区分不同的基因型,适合大量的实验对象;直接测序法交由测序公司测定,样本数量浮动范围较大,费用较高。结论:Taqman-MGB探针法适用于GITRL基因rs2236876A/G基因分型,PCR-RFLP法适用于基因分型结果的验证,直接测序法适用于鉴定有争议的PCR-RFLP法的验证结果。

Objective: Comparing three single nucleotide polymorphism( SNP) genotyping methods,to choose an accurate and effective SNP genotyping method for rs2236876 A / G of GITRL gene. Methods: SNP genotyping for rs2236876 A / G of GITRL were performed and compared by Taqman-MGB probe,the PCRRLFP and direct sequencing. Results: PCR-RLFP showed the genotyping result by electrophoresis,which was suitable for a small number of experimental subjects. Taqman-MGB probe showed the genotyping result by fluorescence,which was suitable for a large number of experimental subjects. Direct sequencing method was operated by the sequencing company which was expensive. Conclusion: For rs2236876 A / G of GITRL gene,Taqman-MGB probe method could be chosen for genotyping,PCR-RFLP method confirmed the genotyping results and,at the same time,the direct sequencing finally validated the result of PCR-RFLP.