Purification of supercoiled plasmid DNA using chromatographic processes
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Updated: Oct 24, 2026

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
Published on: January 5, 2011
G N Ferreira1, J M Cabral, D M Prazeres
1Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal.
Abstract:
The interest in purifying injectable-grade plasmid DNA has increased with the development of gene therapy and DNA vaccination technologies. In this paper we develop a method for purifying a 4.8 kb plasmid based on chromatographic processes. An NaCl gradient was optimized on a Q Sepharose column and plasmid was eluted at 800-820 mM NaCl in a broad peak. Supercoiled plasmid was isolated after a final Sepharcryl S1000 SF gel filtration step. Final plasmid preparation was depleted of proteins and RNA, as revealed by the BCA assay and 1% agarose gel electrophoresis.
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