Open AccessA RAPID PLASMID PREPARATION METHOD BY THE DIRECT BOILING OF ESCHERICHIA COLI CELLS *
Author(s) -
OUYANG LAN,
QIN LIANG,
XU ZHIWU,
HE JIANGUO,
LIU QIUYUN
Publication year - 2008
Publication title -
journal of rapid methods & automation in microbiology
Language(s) - English
Resource type - Journals
eISSN - 1745-4581
pISSN - 1060-3999
DOI - 10.1111/j.1745-4581.2008.00113.x
Subject(s) - plasmid , escherichia coli , plasmid preparation , transformation (genetics) , cloning (programming) , dna , polymerase chain reaction , chemistry , microbiology and biotechnology , chromatography , biology , boiling , ethanol precipitation , extraction (chemistry) , biochemistry , gene , computer science , pbr322 , organic chemistry , programming language
A 10 min plasmid extraction method by the direct boiling of Escherichia coli cells, which avoids isopropanol/ethanol precipitation and drying steps used in standard plasmid preparations, is described. Plasmid DNA thus isolated can be used for restriction characterization, chemical transformation, polymerase chain reaction (PCR) amplification as well as cloning purposes.PRACTICAL APPLICATIONS This method requires minimum experimental setup, and plasmid DNA prepared can be subjected to restriction digestions without visible degradation. It will enable researchers to process hundreds of cell cultures in a single day. Terrific Broth can be adopted for high cell density growth to yield more DNA with our protocol.