【摘要】 目的建立简便易行、经济适用的MGMT基因4个单核苷酸多态性位点的检测方法。方法应用碱基错配创造酶切位点原理设计引物,通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术判断MGMT基因四个SNP位点多态性。结果设计一对引物PCR扩增含MGMT84多态位点的DNA片段,另一对引物PCR扩增含MGMT基因143、160、178三个多态位点的DNA片段,使用4种限制性内切酶分别酶切判断4个位点多态性,PCR和酶切效果均较好。结论应用创造酶切位点PCR-RFLP原理建立的MGMT四个多态位点的检测方法具备简便、经济、快速的特点。更多还原

【Abstract】 Objective To develop a proper assay for identifying single nucleotide polymorphisms(SNPs) of the MGMT gene. Methods PCR primers were designed by create restriction site(CRS) method, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to identify four SNPs in MGMT gene. Results By PCR, one primer pair yielded target products containing MGMT84 SNP site, and the other primer pair yielded target products containing MGMT143,160,178 SNP sites. Four restriction enzymes were adopted to identify the four SNPs, respectively. The effects of PCR and RFLP were good. Conclusion The methods for four SNPs of MGMT determinated by CRS-PCR-RFLP theory could be facility, economy, and rapidness.更多还原