QIAprep Spin Miniprep Kit
For purification of up to 20 μg molecular biology grade plasmid DNA
The QIAprep Spin Miniprep Kit is designed for isolation of up to 20 μg high-purity plasmid or cosmid DNA for use in routine molecular biology applications, including fluorescent and radioactive sequencing and cloning. Even higher yields (up to 30 μg) can be achieved using the High-Yield Supplementary Protocol. The QIAprep Spin Miniprep Kit can be automated on the QIAcube.
For optimal results it is recommended to use this product together with QIAvac 24 Plus.
The QIAprep Spin Miniprep Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
The QIAprep Spin Miniprep Kit enables purification of up to 20 μg molecular biology grade plasmid DNA or cosmid DNA for use in routine molecular biology applications such as PCR, sequencing and cloning. Versatile QIAprep 2.0 Spin Columns can be used either in microcentrifuges, on vacuum manifolds, or in the QIAcube (see figures "QIAprep 2.0 Spin Column handling options A, B, and C"). The vacuum procedure provides simplified handling and faster sample processing. QIAprep 2.0 Spin Columns can be vacuum processed using the QIAvac 24 Plus or any other commercial manifold with luer connectors. The QIAprep Spin Miniprep Kit can now also be fully automated on the QIAcube.
Purified DNA can be used in restriction digestion (see figure "Complete digestion with various restriction enzymes").
Principle
QIAprep 2.0 Spin Columns contain a unique silica membrane that binds up to 20 μg DNA in the presence of a high concentration of chaotropic salt, and allows elution in a small volume of low-salt buffer. QIAprep membrane technology eliminates time consuming phenol–chloroform extraction and alcohol precipitation, as well as the problems and inconvenience associated with loose resins and slurries. High-purity plasmid DNA eluted from QIAprep 2.0 Spin Columns is immediately ready to use – there is no need to precipitate, concentrate, or desalt.
To enable faster and more convenient sample processing and analysis, gel loading dye is provided in the kit. GelPilot Loading Dye contains 3 tracking dyes (xylene cyanol, bromophenol blue, and orange G) to facilitate the optimization of agarose gel run time and prevent smaller DNA fragments migrating too far (see figure "GelPilot Loading Dye").
Procedure
Plasmid purification using QIAprep Kits follows a simple bind-wash-elute procedure (see flowchart "The QIAprep procedure"). First, bacterial cultures are lysed and the lysates are cleared by centrifugation. The cleared lysates are then applied to the QIAprep 2.0 module where plasmid DNA adsorbs to the silica membrane. Impurities are washed away and pure DNA is eluted in a small volume of elution buffer or water.
In addition to plasmid purification from Escherichia coli, QIAprep Kits can be used to purify plasmid DNA from Saccharomyces cerevisiae, Bacillus subtilis, and Agrobacterium tumefaciens. Contact QIAGEN Technical Services or your local distributor for protocols for these applications.
Applications
The QIAprep Miniprep Kits provides reproducible yields of high-purity DNA suitable for use in most applications, including:
Customers who bought these products also bought
|
