论文部分内容阅读
目的 建立一种快速、简便、敏感检测载脂蛋白E(ApoE)基因突变的PCR -SSCP方法。方法 提取人全血基因组DNA ,采用聚合酶链式反应 (PCR)扩增ApoE基因目的片段 ,用甲酰胺变性液进行变性后行 8%聚丙烯酰胺凝胶电泳后 ,对凝胶进行银染 ,然后进行SSCP分析。结果 PCR -SSCP银染图谱显示对ApoE基因双链有很好的分离 ,可观察到对应于不同基因型的不同带型。结论 建立了一种灵敏、准确进行ApoE基因突变分析的SSCP方法
Objective To establish a rapid, simple and sensitive PCR-SSCP method for detecting the mutation of apolipoprotein E (ApoE) gene. Methods Genomic DNA of human whole blood was extracted. ApoE gene fragment was amplified by polymerase chain reaction (PCR), denatured with formamide denatured solution and then subjected to 8% polyacrylamide gel electrophoresis. The gel was stained with silver, Then SSCP analysis. Results PCR-SSCP silver staining showed that the ApoE gene double-stranded DNA was well separated and different bands corresponding to different genotypes were observed. Conclusion A sensitive and accurate SSCP method for ApoE gene mutation was established