TITLE:
A simplified protocol for the semi-large scale recovery of plasmids from Escherichia coli grown on agar plates
AUTHORS:
Masahiro Sato, Eri Akasaka, Issei Saitoh, Masato Ohtsuka, Shingo Nakamura, Takayuki Sakurai, Satoshi Watanabe
KEYWORDS:
Agar Plate; DNA Purification; E. coli; Gene Engineering; Plasmid
JOURNAL NAME:
Journal of Biomedical Science and Engineering,
Vol.5 No.7,
July
3,
2012
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.