一种改良质粒DNA提取方法在实验课中的应用

A Modified Method for Isolation of Plasmid DNA in Experimental Class

改良碱裂解质粒DNA提取方法,将溶液Ⅰ、Ⅱ、Ⅲ、无水乙醇的反应时间分别缩短为0、0、10、0 min;用针式滤器除去残余蛋白质,替代了传统的酚/氯仿抽提方式,缩短DNA提取时间,使其在1 h内完成,并减少有毒物质对人体造成的伤害。获得的质粒DNA质量好、产率高,能满足常规分子生物学实验的要求。

Modifications were made on the preparation of plasmid DNA by alkaline lysis, the reaction time of solutionI, solution Ⅱ , solution Ⅲ and Alcohol was 0 min, 0 min, 10 min, 0 min respectively; residual protein was wipped off by the filter, which instead of phenol and chloroform extraction. Thus the time of isolation of plasmid DNA was shortened within 1 hour, and reducing the injury of poisonous substance on the human body. The experiments demonstrated that the improved method generated good quality plasmid DNA with high yield, and it could meet the standards for many common molecular biology experiments.