For ultrafast purification of up to 750 µg transfection-grade plasmid or cosmid DNA
Less than 60 minutes prep time
Lysate clearing and isopropanol precipitation without centrifugation
No risk of DNA pellet loss during precipitation
Up to 750 µg yield of high-copy plasmid DNA
LyseBlue for optimum lysis and maximum DNA yield
Large-scale plasmid preparation without vacuum manifold
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 are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
QIAprecipitator module.
The unique 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.
QIAfilter Cartridge.
QIAfilter Cartridges 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.
Performance
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 µl – 250 ml or 50 µl – 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.
Principle
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 Maxi Kit
HiSpeed Plasmid Midi Kit
Applications
Transfection, cloning sequencing, gene silencing
Transfection, cloning sequencing, gene silencing
Culture volume/starting material
150 µl – 250 ml culture volume
50 µl – 150 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
60 min
45 min
Yield
<750 µg
<200 µg
Procedure
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.
Applications
DNA purified with HiSpeed Plasmid Kits yield excellent results in all applications, from cloning and sequencing to transfection and plasmid-mediated gene silencing.
