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目的 :建立一种准确、简便和经济的适用于大范围测定载脂蛋白 E基因型的方法。方法 :自外周血中采用 TKM法提取人基因组 DNA,PCR- RFLP确定载脂蛋白 E的基因型 ,硝酸银染色聚丙烯酰胺凝胶 ,若银染效果不佳可用硝酸脱色后重染。结果 :TKM法获得 DNA在产量和质量上均能满足 PCR需求 ;电泳凝胶经银染可得清晰的 DNA特异带 ,据此能准确确定样品载脂蛋白 E的基因型 ;用硝酸溶液脱色后的凝胶经重染仍可得到清晰的图谱。结论 :改进的载脂蛋白E基因型测定方法不仅操作简便、快速 ,而且更经济、更省时 ,适于大范围开展载脂蛋白 E基因型与相关疾病关系的研究
Objective: To establish an accurate, simple and economical method for the determination of apolipoprotein E genotype in a wide range. Methods: Human genomic DNA was extracted from peripheral blood by TKM method. The genotype of apolipoprotein E was determined by PCR-RFLP. Polyacrylamide gel was stained with silver nitrate. If the silver staining was not effective, it could be decontaminated by nitric acid. Results: The yield and quality of DNA obtained by TKM method could meet the requirement of PCR. The gel electrophoresis gel could obtain a clear DNA specific band by silver staining, which could accurately determine the apolipoprotein E genotype of the sample. After decolorization with nitric acid solution The gel is still dyed by a clear map. CONCLUSIONS: The improved apolipoprotein E genotyping assay is not only simple, rapid, economical, and time-saving, but also suitable for the study of the relationship between apolipoprotein E genotype and related diseases in a wide range