QIAquick PCR Purification Kit
For purification of up to 10 μg PCR products, 100 bp to 10 kb
The QIAquick PCR Purification Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of PCR products >100 bp. DNA of up to 10 kb is purified using a simple and fast bind-wash-elute procedure and an elution volume of 30–50 μl. An optional 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.
The QIAquick PCR Purification Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
The QIAquick PCR Purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, and other impurities from DNA samples (see figure "Complete primer removal after PCR"). Using a microcentrifuge or vacuum manifold, DNA ranging from 100 bp to 10 kb is purified.
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 "Complete primer removal after PCR"). 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 dyeTo 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"). Binding buffer is added directly to the PCR sample or other enzymatic reaction, and the mixture is applied to the QIAquick spin column. The binding buffer contains a pH indicator, allowing easy determination of the optimal pH for DNA binding (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. HandlingQIAquick 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 QIAvac 24 Plus with QIAvac Luer Adapters. The QIAquick PCR Purification 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, microarray analysis, ligation and transformation, restriction digestion, labeling, microinjection, PCR, and in vitro transcription.
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