Description
QIAfilter Plasmid Kits provide anion-exchange-based plasmid purification with bacterial lysate clearing by filtration. Purified DNA is equivalent to that obtained by 2 x CsCl gradient centrifugation and is suitable for transfection-grade applications. The QIAfilter Plasmid Mega Kit (cat. no. 12281) and the QIAfilter Plasmid Giga Kit (cat. no. 12291) will be discontinued on June 30, 2021. They remain available as long as stocks last. The QIAGEN Plasmid Mega Kit (cat. no. 12181) and the QIAGEN Plasmid Giga Kit (cat. no. 12191) can be used with the QIAfilter Mega-Giga Cartridges (cat. no. 19781) as an optional protocol step for rapid clearing of bacterial lysates by filtration instead of centrifugation. Visit the QIAGEN plasmid kits web page to learn more.
The QIAfilter Plasmid Kits combines QIAfilter Cartridges for rapid clearing of bacterial lysates by filtration instead of centrifugation, with proven QIAGEN-tips containing anion-exchange resin for efficient plasmid purification. Yields of up to 500 µg (maxi), and 100 µg (midi) transfection-grade, high-copy plasmid DNA are achieved from culture (culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium).
QIAfilter Cartridges, provided in QIAfilter, HiSpeed, and EndoFree Plasmid Kits, are special filter units designed to replace centrifugation following alkaline lysis of bacterial cells. QIAfilter Cartridges completely remove SDS precipitates and clear bacterial lysates in a fraction of the time needed for centrifugation, reducing plasmid-purification time by up to 1 hour. As an optional protocol step for rapid clearing of bacterial lysates by filtration instead of centrifugation, you can use the QIAfilter Mega-Giga Cartridges (cat. no. 19781) with the QIAGEN Plasmid Mega Kit (cat. no. 12181) and the QIAGEN Plasmid Giga Kit (cat. no. 12191). Visit the QIAGEN plasmid kits web page to learn more. QIAfilter Maxi and Midi Cartridges use a syringe format and lysates are cleared rapidly by pushing the liquid through the filter. The unique anion-exchange resin in QIAGEN-tips is developed exclusively for the purification of nucleic acids. Its exceptional separation properties result in DNA purity equivalent or superior to that obtained by two successive rounds of CsCl gradient centrifugation. Prepacked QIAGEN-tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment. Specifications Features QIAfilter Plasmid Maxi Kit QIAfilter Plasmid Midi Kit Applications Transfection, cloning, sequencing, etc. Transfection, cloning, sequencing, etc. Culture volume/starting material 100–250 ml culture volume 25–50 ml culture volume Plasmid type High-copy, low-copy, cosmid DNA High-copy, low-copy, cosmid DNA Processing Manual (filtration) Manual (filtration) Sample per run 1 sample per run 1 sample per run Technology Anion-exchange technology Anion-exchange technology Time per run 120 min 110 min Yield <500 µg <100 µg
Neutralized bacterial lysates are incubated directly in the QIAfilter Cartridge and cleared rapidly by filtration. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and ultrapure plasmid DNA is eluted in high-salt buffer (see flowchart "QIAGEN Plasmid Kit procedures"). The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
