28506-QIAquick PCR & Gel Cleanup Kit (100)- Qiagen

Product Details

The QIAquick Gel Extraction Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of DNA fragments from gels (up to 400 mg slices) or enzymatic reactions. DNA ranging from 70 bp to 10 kb is purified using a simple and fast bind-wash-elute procedure and an elution volume of 30–50 μl. An integrated pH indicator allows easy determination of the optimal pH for DNA binding to the spin column. The procedure can be fully automated on the QIAcube Connect.

For optimal results it is recommended to use this product together with QIAvac 24 Plus.

Performance

The QIAquick Gel Extraction Kit enables removal of nucleotides, enzymes, salts, agarose, ethidium bromide, and other impurities from samples, ensuring up to 80% recovery of DNA (see figure ” High recoveries from gels”). Using a microcentrifuge or vacuum manifold, DNA ranging from 70 bp to 10 kb is purified from 1–24 samples. Purified DNA can be used, for example, in sequencing (see figure ” Reliable sequencing after gel extraction”). DNA fragments smaller than 70 bp or larger than 10 kb should be extracted with the QIAEX II Gel Extraction System.

Principle

QIAquick Kits contain a silica membrane assembly for binding of DNA in high-salt buffer and elution with low-salt buffer or water. The purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from DNA samples (see figure ” High recoveries from gels”). Silica-membrane technology eliminates the problems and inconvenience associated with loose resins and slurries. Specialized binding buffers are optimized for specific applications and promote selective adsorption of DNA molecules within particular size ranges.

Gel loading dye

To enable faster and more convenient sample processing and analysis, gel loading dye is provided. GelPilot Loading Dye contains three 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

The QIAquick system uses a simple bind-wash-elute procedure (see flowchart ” QIAquick and MinElute procedure”). Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the QIAquick spin column (see figure ” pH Indicator Dye”). Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

Handling

QIAquick spin columns are designed to provide two convenient handling options. The spin columns fit into a conventional table-top microcentrifuge or onto any vacuum manifold with luer connectors, such as the QIAvac 24 Plus with QIAvac Luer Adapters. The QIAquick Gel Extraction Kit, in addition to other QIAGEN spin-column-based kits, can be fully automated on the QIAcube, enabling increased productivity and standardization of results (see figures “Spin column handling options A, B, C, D, and E”).

Applications

DNA fragments purified with the QIAquick system are ready for direct use in all applications, including sequencing, ligation and transformation, restriction digestion, labeling, microinjection, PCR, and in vitro transcription.