QIAprep Spin Miniprep Kit

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.

 

QIAprep Spin Miniprep Kit specifications

Format	Spin columns
Purification module	QIAprep 2.0 Spin Columns
Throughput	1–24 samples
Preparation time	24 minipreps in 30 minutes
Equipment required	Microcentrifuge or vacuum manifold; fully automatable using the QIAcube
Lysate clearing	Centrifugation
Capacity of column reservoir	800 µl
Minimum elution buffer volume	50 µl
Culture volume for high-copy plasmids	1–5 ml
Culture volume for low-copy plasmids/cosmids	1–10 ml

Purified DNA can be used in restriction digestion (see figure "Complete digestion with various restriction enzymes"). 

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").

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.

The QIAprep Miniprep Kits provides reproducible yields of high-purity DNA suitable for use in most applications, including:

• PCR
• Restriction digestion
• Ligation and transformation
• Sequencing
• Screening