Kurien B T, Scofield R H
Oklahoma Medical Research Foundation, University of Oklahoma Health Sciences Center, Oklahoma City, USA.
Biotechniques. 1995 Jun;18(6):1023-6.
A moderately efficient and quick method of bacterial colony transformation is described. Plasmid DNA was added to bacteria suspended in a solution of polyethylene glycol/calcium chloride (PEG/CaCl2). After a brief incubation and heat shock, the cells were directly plated. Transformation efficiencies up to 8.6 +/- 1.28 x 10(6) transformants per microgram of pUC18 were obtained. We have found that the reverse of the transformation process could also take place. Suspending a bacterial pellet harboring the plasmid of interest in PEG/CaCl2 results in the release of the plasmid DNA, and thus indirectly lends support to the transformation process.
描述了一种中等效率且快速的细菌菌落转化方法。将质粒DNA添加到悬浮于聚乙二醇/氯化钙(PEG/CaCl₂)溶液中的细菌中。经过短暂孵育和热休克后,细胞直接铺板。获得了高达每微克pUC18 8.6±1.28×10⁶个转化子的转化效率。我们发现转化过程的逆过程也可能发生。将含有感兴趣质粒的细菌沉淀悬浮于PEG/CaCl₂中会导致质粒DNA的释放,从而间接支持了转化过程。