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

Masahiro Sato; Eri Akasaka; Issei Saitoh; Masato Ohtsuka; Shingo Nakamura; Takayuki Sakurai; Satoshi Watanabe

doi:10.4236/jbise.2012.57051

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JBiSE> Vol.5 No.7, July 2012

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

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DOI: 10.4236/jbise.2012.57051 3,918 Downloads 6,514 Views Citations

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Masahiro Sato, Eri Akasaka, Issei Saitoh, Masato Ohtsuka, Shingo Nakamura, Takayuki Sakurai, Satoshi Watanabe

Affiliation(s)

Animal Genome Research Unit, Division of Animal Science, National Institute of Agrobiological Sciences, Ibaraki, Japan.
Department of Organ Regeneration, Graduate School of Medicine, Shinshu University, Nagano, Japan.
Department of Pediatric Dentistry, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
Department of Surgery II, National Defense Medical College, Saitama, Japan.
Division of Basic Molecular Science and Molecular Medicine, School of Medicine, Tokai University, Kanagawa, Japan.
Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan.

ABSTRACT

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.

KEYWORDS

Agar Plate; DNA Purification; E. coli; Gene Engineering; Plasmid

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

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.

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