单链构象多态性-毛细管电泳分析中样品纯化研究

Effect of Sample Purification on Single-strand Conformation Polymorphism Analysis by Capillary Electrophoresis

聚合酶链反应(PCR)样品中的Cl^-、引物等小分子对单链构象多态性-毛细管电泳分析有重要影响。本文用毛细管电泳-间接紫外法测定了PCR样品中的Cl^-浓度,并对乙醇沉淀法和试剂盒纯化法用于PCR样品的纯化效果进行比较,在此基础上研究了Cl^-浓度、引物及其二聚体在SSCP-CE分析中的影响。结果表明,它们主要影响DNA进样量而对分离效率影响不大。

Chloride and primers present in the PCR sample have an effect on single-strand conformation polymorphism analysis by capillary electrophoresis using electrokinetic injection. In this paper, the concentration of chloride ion in the PCR sample was quantitatively determined by a CE-indirect UV method. The ethanol precipitation method and PCR sample purified kit for purified PCR samples were compared. The effects of chloride ion concentration and primers on SSCP-CE analysis were investigated. It was found that the amount of DNA fragments injected into the capillary column decreased with the increase of chloride concentration, while the separation efficiency of DNA fragments did not change.