[目的]对常规碱裂解法进行改进.[方法]分别改变溶液Ⅰ、Ⅱ、Ⅲ及无水乙醇的反应时间,少量提取质粒DNA.根据产率和纯度,得出各溶液的最佳反应时间,以紫外分光光度法、琼脂糖凝胶电泳结果验证提取效果.[结果]将常规碱裂法中溶液Ⅰ、Ⅱ、Ⅲ及无水乙醇的反应时间由原来的5、5、5、15 min分别缩短至0、1、1、5 min,纯度达1.8~1.9,产率达180.0~248.0 μg/ml.[结论]改良法提取质粒DNA,总提取时间缩短了23 min.%[Objective] The alkaline lysis method was improved. [Method] To extract a small amount of plasmid DNA, the reaction time of solution Ⅰ,Ⅱ , Ⅲ and ethanol was changed,based on the yield and purity,the optimum reaction time was obtained. The purification efficiency in the experiment was to be tested by UV spectrophotometer examination and agarose gel electrophoresis. [ Result ] Decrease the reaction time of solution Ⅰ , Ⅱ , Ⅲ and ethanol from 5,5,5,15 min to 0,1,1,5 min respectively, the purity was between 1. 8 and 1.9, the yield was between 180.00 and 248.0 μg/ml. [Conclusion] The overall DNA extraction time using improved alkaline lysis method was reduced by 23 minutes.
展开▼