大肠杆菌感受态细胞的少量制备方法及转化条件研究

Study on the Mini-preparation Method and Transformation Conditions for the Competent Cells of Escherichia coli

[目的]提高大肠杆菌感受态细胞的转化效率。[方法]根据普通实验室的试验条件,总结出1种少量制备大肠杆菌感受态细胞并高效率转化质粒的方法。[结果]利用该方法少量制备大肠杆菌感受态细胞,在OD600nm值为0.3～0.6时均可达到高转化率的效果。与常用的大量制备法相比,该方法的转化率提高100～300倍。[结论]该方法简单方便、重复性好、转化效率高、成本低,能够满足基因库的构建、分子克隆等研究的要求,是从事基因克隆研究方面的科研人员的良好选择。

[Objective] The aim of the research was to increase the transformation efficiency of the competent cells of Escherichia coll. [Method] According to the test conditions in the common laboratory, a kind of method for preparing small amount of competent cells of E. coli and transforming the plasmid effectively was summarized. [Result] The competent cells of E. coli was mini-prepared by this method and the effect with higher transformation efficiency could all be reached at OD600nm, value of 0.3 -0.6. Compared with the commonly -used mass preparation method, the transformation efficiency by this method was increased by 100-300 times. [Conclusion] This method was simple and convenient, with good repeatability, high transformation efficiency and low cost. It could meet the demands of such researches as the construction of gene bank and molecular cloning and it was good choice for the scientific researchers who engaged in the research of gene cloning.