摘要
目的:聚合酶链反应-单链构象多态性(PCR-SSCP)是一种快速、简便、较灵敏的检测基因变异的方法,但其影响因素很多,本文对凝胶浓度、上样量、PCR产物稀释倍数、甘油浓度、电压、电泳温度几个重要的因素进行探讨。方法:通过应用PCR-SSCP方法结合核苷酸序列分析检测成都汉族群体98个个体人类补体第8成分(C8A)基因型频率分布。结果:凝胶浓度为8%,PCR产物稀释4倍,上样量为4μl,不加甘油,电压为200V,恒温电泳时,SSCP分析可得到满意的结果。结论:PCR-SSCP分析对不同的研究对象,其分析参数不同,需要对其分析条件进行最大优化,才能获得实验的成功。
Objective： To study the experimental conditions of polymerase chain reaction-single strand conformation polymorphism （PCR-SSCP） analysis. Methods： The DNA polymorphism of the human eighth complement component （C8A） and its genotype frequency distribution in Chengdu Han population were studied with PCR-SSCP analysis followed by DNA sequencing in different experimental conditions and compared. Results： The optional conditions were as followed： the gel-concentration is 8 %, the dilution of the PCR products is 1 ： 4, the sample quantity of the PCR products applied in electrophoresis is 4μl, no glycerol in gels, during electrophoresis, the voltage is 200V, with stable temperature. Conclusion： Because different DNA fragments entail their own optional running conditions, PCR-SSCP analysis is capable of detecting different samples only under different optimized conditions.
出处
《泸州医学院学报》
2005年第5期391-393,共3页
Journal of Luzhou Medical College
基金
四川省学术带头人培养基金资助项目(4200313)
