摘要
为提高单核苷酸多态性检测的通量,引入多重嵌合引物PCR和毛细管电泳对四引物扩增受阻突变体系PCR进行改进.针对乳腺癌位点rs4784227(C>T),rs1219648(G>A)和rs3803662(T>C)设计特异性嵌合引物,经一次PCR扩增后,通过毛细管电泳分析产物长度,同时确定3个位点的基因型.70份全血和口腔拭子样本,电泳结果均与测序一致,实现成功分型.本方法仅需一次PCR和一次毛细管电泳即可获得3个位点的分型结果,操作简单、快速准确.
We describe a multiplex tetra-primer amplification refractory mutation system PCR（M-T-ARMS PCR）for detecting three single nucleotide polymorphisms associated with breast cancer,including rs4784227（C〉T）,rs12196489（G〉A） and rs3803662（T〉C）.Three sets of chimeric primers consisting of a specific sequence fused to a universal sequence at the 5’end were used in one single PCR reaction,followed by a capillary electrophoresis to separate PCR products and genotyping.We applied this method to test 70 samples from whole blood and mouth swab specimens to demonstrate the performance.All mutations were accurately detected and results were confirmed by direct DNA sequencing.M-T-ARMS PCR coupled with capillary electrophoresis is a simple and accurate assay that allows the simultaneous detection of three mutations in a single tube.
出处
《中国科学：生命科学》
CSCD
北大核心
2012年第2期143-147,共5页
Scientia Sinica(Vitae)
关键词
单核苷酸多态分型
四引物扩增受阻突变体系PCR
多重PCR
毛细管电泳
single nucleotide polymorphisms； genotyping； tetra-primer amplification refractory mutation system PCR； multiplex PCR； capillary electrophoresis
