阿胶DNA提取方法优化

Improvement of Extraction Method of DNA from Asini Corii Colla

目的:优选阿胶DNA的提取方法和效率,为其他高蛋白含量产品中DNA的提取提供参考。方法:采用十二烷基苯磺酸钠(SDS)法和十六烷基三甲基溴化铵(CTAB)法提取阿胶DNA,针对阿胶产品的自身特性,优选SDS法中蛋白酶K作用时间和蛋白洗脱步骤。结果:在同等条件下,SDS法较CTAB法对阿胶DNA的平均提取量高0.022 g·L-1。优化的SDS法为蛋白酶K作用时间1.5 h,使用酚-三氯甲烷-异戊醇洗脱阿胶中蛋白质,重复3次;阿胶DNA的纯度较原工艺提高了48.3%。结论:采用SDS法对阿胶中蛋白的清除作用较CTAB法效率高,优化的SDS法对阿胶DNA的提取适用性较强,为阿胶中动物来源分子生物学检测提供条件。

Objective：To improve extraction efficiency of DNA in Asini Corii Colla and provide a reference for extracting DNA in other products containing high protein content.Method; DNA in Asini Corii Colla was extracted by SDS method and CTAB method,according to their own characteristics of Asini Corii Colla products,reaction time of proteinase K and protein elution step in SDS method were optimized.Result：Under the same conditions,average extraction capacity of DNA in Asini Corii Colla by SDS was higher than CTAB,whose numerical difference was 0.022 g ·L-1 Optimized SDS method was as following：reaction time of proteinase K 1.5 h,eluted protein in Asini Corii Colla by phenol-chloroform-isoamyl alcohol for three times,purity of DNA in Asini Corii Colla increased 48.3％ by comparing with the original process.Conclusion：Scavenging effects on protein in Asini Corii Colla by SDS method is higher than CTAB,this optimized SDS method is suitable for extraction of DNA in Asini Corii Colla,it provides conditions for biological detection of animal derived molecules from Asini Corii Colla.