Purification of plasmid DNA using a multicompartment electrolyser separated by ultrafilter membranes |
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Authors: | Zhijun Wang Guowei Le Yonghui Shi Grzegorz Węgrzyn |
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Affiliation: | (1) Animal Science Research Institute, Southern Yangtze University, Wuxi, 214036, People's Republic of China;(2) Department of Molecular Biology, University of Gdánsk, Kladki 24, 80-822 Gdánsk, Poland |
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Abstract: | A simple, scalable method for purification of plasmid DNA is described. Plasmid DNA was released from Escherichia coli JM109 by lysis (1% SDS, 0.2 M NaOH). Then a neutralization solution (3 M sodium acetate buffer, pH 4.8) was added to precipitate genomic DNA and protein. After the clarification of the lysate, the supernatant was placed in a multicompartment electrolyser separated by ultrafilter membranes to remove the remaining contamination (RNA, genomic DNA and protein). A recovery of 75%±2% of total plasmid DNA was obtained after 60 min electrophoresis with a field strength of 8 V cm–1 using cells at 30 g l–1 (quantified by dry cell weight). Genomic DNA, RNA and protein were undetectable in the purified plasmid DNA solution. |
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Keywords: | DNA vaccine electroseparation gene therapy multicompartment electrolyser plasmid purification |
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