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聚乙二醇介导的细菌菌落转化

Polyethylene glycol-mediated bacterial colony transformation.

作者信息

Kurien B T, Scofield R H

机构信息

Oklahoma Medical Research Foundation, University of Oklahoma Health Sciences Center, Oklahoma City, USA.

出版信息

Biotechniques. 1995 Jun;18(6):1023-6.

PMID:7546702
Abstract

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.

摘要

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。

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