For ultrafast purification of up to 200 µg transfection-grade plasmid or cosmid DNA

  • Less than 45 minutes prep time
  • Lysate clearing and isopropanol precipitation without centrifugation
  • No risk of DNA pellet loss during precipitation
  • Up to 200 µg yield of high-copy plasmid DNA
  • LyseBlue for optimum lysis and maximum DNA yield

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.

Quantity Product Cat. no. Price
 
 
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  varies
Quantity Product Cat. no. Price Sum
 
HiSpeed Plasmid Midi Kit (25)
25 HiSpeed Midi Tips, 25 QIAfilter Midi Cartridges, 25 QIAprecipitator Midi Modules plus Syringes, Reagents, Buffers
12643
$320.00
$0.00
The HiSpeed Plasmid Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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QIAGEN Plasmid Kit procedures.|QIAprecipitator module.|QIAfilter Cartridge.|
Neutralized bacterial lysates are incubated directly in the QIAfilter Cartridge and cleared in seconds 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. The DNA is concentrated and desalted without centrifugation using 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 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 provide QIAfilter Cartridges, HiSpeed Tips, and QIAprecipitator modules for fast large-scale plasmid preparation. Syringe-format QIAfilter and QIAprecipitator modules replace centrifugation steps in the classic anion-exchange procedure, making plasmid purification faster and more convenient. Up to 200 µg high-copy plasmid DNA is purified from 50–150 ml culture. (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.

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 yields excellent results in all applications, from cloning and sequencing to transfection and plasmid-mediated gene silencing.

Feature
Specifications
Applications Transfection, cloning, sequencing, gene silencing
Culture volume/starting material 50 µl – 150 ml culture volume
Plasmid type High-copy, cosmid DNA
Processing Manual (filtration)
Samples per run; throughput 1 sample per run
Technology Anion-exchange technology
Time per run or prep per run <45 min
Yield <200 µg

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