QIAquick Nucleotide Removal Kit

• Up to 95% recovery of ready-to-use DNA
• Fast and convenient procedure
• Cleanup of DNA up to 10 kb in three easy steps
• Gel loading dye for convenient sample analysis

The QIAquick Nucleotide Removal Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of oligonucleotides and DNA. Unincorporated nucleotides, salts, and other contaminants are removed and oligonucleotides (>17 nt) and DNA fragments ranging from 40 bp to 10 kb are purified using a simple and fast bind-wash-elute procedure and an elution volume of 30–200 µl. The procedure is fully automated on the QIAcube Connect.

The QIAquick Nucleotide Removal procedure removes nucleotides, enzymes, salts, and other impurities from DNA samples (see figure " Complete removal of nucleotides from labeled oligos". Using a microcentrifuge, 17mer – 10 kb DNA is purified. For sample volumes smaller than 50 µl the DyeEx Spin kit can also be used.

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 removal of nucleotides from labeled oligos"). 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 from migrating too far (see figure " GelPilot Loading Dye").

The QIAquick system uses a simple bind–wash–elute procedure (see flowchart " QIAquick Nucleotide Removal procedure"). Binding buffer is added directly to the sample, and the mixture is applied to the QIAquick spin column. 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. The QIAquick Nucleotide Removal Kit, in addition to other QIAGEN spin-column-based kits, can be fully automated on the QIAcube Connect, enabling increased productivity and standardization of results (see figures "Spin column handling options  A" and " QIAcube Connect").

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, and microinjection.