CaCl_2法质粒转化最佳条件的探讨

The Optimal Condition of Transforming Plasmid into E coli Using Calcium Chloride

ＣａＣｌ２法质粒转化中的一些关键性步骤的实验条件进行了探讨结果表明，ＣａＣｌ２处理对制备感受志受体菌是至关重要的，ＣａＣｌ２处理后的受体菌在４℃放置１６～２４ｈ是必要的．ＣａＣｌ２处理浓度以０．０７５ｍｏｌ·Ｌ－１最佳．为了得到足够的转化菌落，质粒加入量不应低于０．２μｇ．

The authors have inquiried into the optimal experiment conditions of transforming plasmid into E coli using CaC12. The results follows: Fresh competent E coli prepared using CaCl2 is critical and after CaCl2 treatment it is necessary to store the E colt at 4℃ for 16~24h. While 0.025mol. L-1 CaCl2 treatment can prepare competent E coli,but the optimal concentration of CaCl2 is 0.075mol. L-1. To get enough clones,plasmid outlay should not be less than 0.2μg.