摘要
目的探讨多重聚合酶链反应-限制性片断长度多态性(多重PCR-RFLP)技术在单核苷酸多态性检测中的应用。方法应用多重聚合酶链反应(Multiplex)(多重PCR)原理设计人类X射线交错互补修复基因1(XRCC1基因)194、399位点引物,通过PCR-RFLP技术判断XRCC1基因2个SNP位点多态性。结果设计的两对引物可同时PCR扩增分别含2个多态位点的DNA片段,MspI限制性内切酶酶切可判断2位点多态性,PCR和酶切效果均较好。结论多重PCR-RFLP技术可应用于单核苷酸多态位点的检测,并节省时间和费用,提高检测效率。
Objective To explore the strategy for identifying single nucleotide polymorphism（SNP）with technique of multi-PCR by primer design combined with PCR-RFLP.Methods Primer was designed with multi-PCR theory,then using PCR-RFLP to identify 2 SNPs of XRCC1 gene.Results One pair of primers could simultaneously amplify DNA segments containing 2 SNP sites by PCR,while MspI restriction enzyme could identify 2 SNPs,both of them showed good effect.Conclusion It was suggested that Multi-PCR combined with PCR-RFLP can be used to identify SNP,and is good in time and money saving and detect efficiency improving.
出处
《中国工业医学杂志》
CAS
北大核心
2008年第1期3-6,共4页
Chinese Journal of Industrial Medicine
基金
国家自然科学基金(30671740
30070650)
国家973项目(2002CB512909)
复旦大学研究生创新基金资助
关键词
多重PCR
PCR-RFLP
单核苷酸多态性
XRCC1
引物设计
Multiplex polymerase chain reaction（multiplex PCR）
Polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）
Single nucleotide polymorphism（SNP）
X-ray repair cross complementing gene 1（XRCC1）
Primer design
