两种方法制备模板DNA在PCR-SSCP中的应用研究

Research on Application of PCR-SSCP in Preparation of Sample DNA with Two Methods

目的:比较两种方法(DNA试剂盒提取法和FTA卡法)提取的DNA在PCR—SSCP反中的可靠性。方法:用DNA试剂盒从全血中提取DNA和用NaOH方法从FTA卡中提取DNA后,用分光光度计检测两种方法提取DNA的浓度及纯度,进行PCR反应之后,用2%的琼脂糖凝胶电泳检测其质量,接着进行SSCP检测,观测其效果。两种方法提取的样品相同,进行PCR反应和SSCP检测的条件完全一致。结果:用DNA试剂盒提取法和FTA卡法提取的DNA纯度分别为OD260/280=1.817,OD260/280=1.806。48份贵州荷斯坦奶牛DNA后续PCR反应和SSCP的检测结果表明,两种方法提取的DNA用于PCR反应和SSCP检测其效果没有明显差别,成功率为100%。结论:FTA卡结合DNA较稳定,用NaOH法提取DNA效果可靠且比试剂盒方法简便、快捷、经济,值得推广。

Objective：This study tested the reliability of DNA, which using NaOH method extract from FTA card and kit extraction, in PCR reaction, PCR -SSCP reaction. Method：The concentration and purity of DNA, which extracted in two different methods, was tested by spectrophotometer. Then PCR reaction and PCR - SSCP reaction were proceed and the results were checked up. The samples were the same and the two reactions were proceed in same conditions. Result：The purity of DNA were OD260/280 = 1. 817 and OD260/280 = 1. 806,re- spectively. The results of PCR and PCR - SSCP indicated no difference between them and success rate is 100% in all 48 samples. Conclusion：Therefore, the DNA samples adhered properly to FTA card, it is reliable which using NaOH method extract DNA, and more simple and inexpensive. It＇s worth to popularize this method.