鲑鱼精DNA提高人工突变质粒模板中单核苷酸多态性位点识别的特异性及其应用  

Salmon DNA Improve the Identification of the Gene's SNP on Plasmid with Artificial Mutation and Its Application

作　　者：刘二平[1] 韩立影[2] 方琪[2] 秦正红[1] 王进[1] 

机构地区：[1]苏州大学药学院药理学系衰老和神经疾病实验室,江苏苏州215123 [2]苏州大学附属第一医院神经内科,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2011年第3期380-383,399,共5页Suzhou University Journal of Medical Science

基　　金：国家高技术研究发展计划(863计划)项目(2008AA02Z436)

摘　　要：目的探讨等位基因特异性聚合酶链反应(AS-PCR)检测单核苷酸多态性(SNP)的方法,研究鲑鱼精DNA对优化等位基因特异性引物的作用。方法对鲑鱼精DNA设一系列浓度梯度,分别进行AS-PCR后观察电泳结果。结果鲑鱼精DNA能显著提高识别基因SNP位点的特异性,在25μl的质粒检测系统中加入50 ng的鲑鱼精DNA结果最佳。结论鲑鱼精DNA加入到质粒检测系统中可有效测试所设计的检测引物的特异性,达到模拟基因组DNA的目的 。Objective To study the effection of salmon DNA on improving single nucleotide polymorphism（SNP） detection specificity in AS-PCR with plasmid DNA template in order to facilitate allele specific primer optimization.Methods Using SNP-containing plasmind DNA with salmon DNA at va-rious concentrations was performed AS-PCR and SNP detection specificity among them was compared after agarose gel electrophoresis.Results With increase of concentrations,Salmon DNA could significantly enhanced the discrimination power of SNP genotyping by AS-PCR using plasmid DNA.By balancing between enhancing effect on SNP genotyping and reducing effect on PCR efficiency,the best system was achieved in a 25 μl reaction system with 50 ng salmon DNA and 0.01 ng plasmid DNA.Conclusion Salmon DNA can significantly improve SNP discrimination power by AS-PCR with plasmid DNA,which mimics AS-PCR with human genomic DNA as template.

关 键 词：鲑鱼精DNA 单核苷酸多态性 等位基因特异性聚合酶链反应 

分 类 号：R394-33[医药卫生—医学遗传学]