A simplified protocol for the semi-large scale recovery of plasmids from Escherichia coli grown on agar plates英文文献资料.docVIP

J. Biomedical Science and Engineering, 2012, 5, 406-408 JBiSE /10.4236/jbise.2012.57051 Published Online July 2012 (http://www.SciRP.org/journal/jbise/) 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 , 1 2 2 3 4 5 6 Satoshi Watanabe 1 2 3 4 5 6 Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan Department of Pediatric Dentistry, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan Division of Basic Molecular Science and Molecular Medicine, School of Medicine, Tokai University, Tokyo, Japan Department of Surgery II, National Defense Medical College, Tokorozawa, Japan Department of Organ Regeneration, Graduate School of Medicine, Shinshu University, Matsumoto, Japan Animal Genome Research Unit, Division of Animal Science, National Institute of Agrobiological Sciences, Tsukuba, Japan Email: masasato@ms.kagoshima-u.ac.jp Received 9 May 2012; revised 8 June 2012; accepted 28 June 2012 ABSTRACT paratus, a process that takes approximately 40 - 60 min. The collected supernatant is then subjected to further pu- rification by using column [1,2] or density-gradient ul- tracentrifugation of a cesium chloride/ethidium bromide (EtBr) mixture [3]. Growth of E. coli on agar plates is commonly used for the preparation of MiniPrep DNA [3], but rarely used for isolation of plasmids at a semi-large scale level. This procedure does not require the above-mentioned centri- fugation apparatus, but requires ultra-rapid centrifuga- tion equipment, which is generally used for precipitation of pl