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发明公开
CN105274204A 人PLIN1基因rs894160位点多态性检测方法及试剂盒 无效 - 驳回

人PLIN1基因rs894160位点多态性检测方法及试剂盒
摘要：
人PLIN1基因rs894160位点多态性检测方法及试剂盒。该方法，包括：提供待测人基因组DNA；提供扩增人PLIN1基因rs894160位点附近序列的上游引物和下游引物，其中下游引物具有错配碱基G；以所述待测人基因组DNA为模板，利用上游引物和下游引物进行PCR扩增反应以获得含ARGCTT片段或者RGCT片段的扩增产物，其中R为人PLIN1基因rs894160位点上的待定碱基A或G；提供限制性内切酶；利用限制性内切酶对所得扩增产物进行酶切以获得相应的酶切产物；以及根据所得酶切产物来确定人PLIN1基因rs894160位点上的待定碱基R是A还是G。限制性内切酶为能够切开AAGCTT片段与AGGCTT片段之一，或者AGCT片段与GGCT片段之一的限制性内切酶。本发明的测定手段不但快速可靠，而且测定成本大大降低。
摘要（英）：

The invention discloses a method and a kit for determination of human PLIN1 gene rs894160 site polymorphism. The method is as follows: providing to-be-tested human genomic DNA; providing upstream and downstream primers for amplification of sequences nearby human PLIN1 gene rs894160 site, wherein the downstream primer has a mismatched base G; taking the to-be-tested human genomic DNA as a template, using the upstream and downstream primers for PCR amplification reaction to obtain an amplification product containing ARGCTT or RGCT fragment, wherein R is to be confirmed base A or G on the human PLIN1 gene rs894160 site; providing a restriction endonuclease; using the restriction endonuclease for enzyme digestion of the amplification product to obtain a corresponding enzyme digestion product; and according to the resulting enzyme digestion product, determining whether the to be confirmed base R on the human PLIN1 gene rs894160 site is A or G. The restriction endonuclease is restriction endonuclease which only be capable of cutting one of AAGCTT fragment or AGGCTT fragment, or restriction endonuclease which only be capable of cutting one of AGCT fragment or GGCT fragment. The determination method is fast and reliable, and can greatly reduce the determination cost.

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