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An improved system for competent cell preparation and high efficiency plasmid transformation using different Escherichia coli strains Electron. J. Biotechnol.
Tu,Zhiming; He,Guangyuan; Li,Kexiu X.; Chen,Mingjie J.; Chang,Junli; Chen,Ling; Yao,Qing; Liu,Dongping P.; Ye,Huan; Shi,Jiantao; Wu,Xuqian .
This paper describes an efficient bacterial transformation system that was established for the preparation of competent cells, plasmid preparation, and for the storage in bacterial stocks in our laboratory. Using this method, a number of different plasmids have been amplified for further experiments. Competent cells for bacterial transformation were prepared by the calcium chloride method with an optimum concentration of 75 mM. Three different strains of Escherichia coli that were tested are DH5α, TG1 and XL1 blue, and the most efficient strain being XL1 blue. The optimal optical density (OD600) range for competent cell preparation varied for each of the strains investigated, and for XL1 blue it was 0.15-0.45; for TG1 it was 0.2-0.5; and for...
Tipo: Journal article Palavras-chave: Competent cells; E. coli; Plasmid; Storage; Transformation.
Ano: 2005 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000100014
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