Description
HiSpeed Plasmid Kits provide fast, large-scale, anion-exchange-based plasmid DNA preparation without centrifugation. The purified DNA is equivalent to that obtained by 2 x CsCl gradient centrifugation and is suitable for transfection-grade applications.
HiSpeed Plasmid Kits contain QIAfilter Cartridges, HiSpeed Tips, and QIAprecipitator modules for fast large-scale plasmid preparation without vacuum manifold. Syringe-format QIAfilter and QIAprecipitator modules replace centrifugation steps in the classic anion-exchange procedure, making plasmid purification faster and more convenient. Up to 750 µg (maxi) or 200 µg (midi) high-copy plasmid DNA can be purified from 150 ml – 250 ml or 50 ml – 150 ml culture, respectively (culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium). HiSpeed Tip design allows a very high flow rate, permitting DNA binding, washing, and elution steps for plasmid purification to proceed faster. The unique anion-exchange resin in HiSpeed 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.
QIAfilter Cartridges (see figure "QIAfilter Cartridge") are special filter units designed to replace the centrifugation step 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. Prepacked HiSpeed Tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. The unique QIAprecipitator module (see figure "QIAprecipitator module") replaces the centrifugation step traditionally used to collect isopropanol-precipitated DNA following purification. The QIAprecipitator module traps the precipitated DNA, while the isopropanol-buffer mixture flows through. The DNA is then simply eluted from the QIAprecipitator into a microcentrifuge tube with TE buffer or water. This unique module also eliminates the risk of pellet loss, which can occur during decanting of the supernatant following centrifugation. Specifications Features HiSpeed Plasmid Midi Kit HiSpeed Plasmid Maxi Kit Applications Transfection, cloning sequencing, gene silencing Transfection, cloning sequencing, gene silencing Culture volume/starting material 50 µl – 150 ml culture volume 150 µl – 250 ml culture volume Plasmid type High-copy, cosmid DNA High-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 45 min 60 min Yield <200 µg <750 µg
Neutralized bacterial lysates are incubated in the QIAfilter Cartridge and cleared in seconds by filtration. The filtrate is applied to a HiSpeed tip for plasmid DNA purification (see flowchart "QIAGEN Plasmid Kit procedures"). Eluted DNA is mixed with isopropanol and applied to the QIAprecipitator module using the syringe provided. The concentrated and desalted DNA is then eluted from the QIAprecipitator directly into a microcentrifuge tube with TE buffer or water.
