A simplified protocol for the semi-large scale recovery of plasmids from Escherichia coli grown on agar plates

DOI: 10.4236/jbise.2012.57051   PDF   HTML   XML   4,092 Downloads   6,700 Views   Citations


Semi-large scale liquid cultivation of transformed Escherichia coli (E. coli) in medium (100-200 ml) has been widely used for the acquisition of relatively large amounts of plasmid DNA (50-300 μg). However, this method requires an expensive high-speed centrifugation apparatus to precipitate E. coli before lysis, which is both laborious and time-consuming. Here, we demonstrate a method for agar plate-based cultivation of bacteria that does not employ a high-speed centrifugation apparatus. This procedure proves to be simple and reproducible, yielding an average of 82 μg of plasmid DNA per experiment. It may therefore be valuable for cloning/transfection experiments under limited financial backgrounds.

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Sato, M. , Akasaka, E. , Saitoh, I. , Ohtsuka, M. , Nakamura, S. , Sakurai, T. and Watanabe, S. (2012) A simplified protocol for the semi-large scale recovery of plasmids from Escherichia coli grown on agar plates. Journal of Biomedical Science and Engineering, 5, 406-408. doi: 10.4236/jbise.2012.57051.

Conflicts of Interest

The authors declare no conflicts of interest.


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