QIAquick Nucleotide Removal Kit

For up to 10 μg oligonucleotide (17–40mers) and DNA (40 bp to 10 kb) cleanup from enzymatic reactions
  • 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.

Quantity Product Cat. no. Price
 
 
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Quantity Product Cat. no. Price Sum
 
QIAquick Nucleotide Removal Kit (50)
50 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)
28304
$108.00
$0.00
 
QIAquick Nucleotide Removal Kit (250)
250 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)
28306
$514.00
$0.00
The QIAquick Nucleotide Removal Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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QIAquick Nucleotide Removal procedure.|Complete removal of nucleotides from labeled oligos.|GelPilot Loading Dye.|Spin column handling options — B.|Spin column handling options — A.|
|Polyacrylamide gel analysis of radioactive labeling reactions before (b) and after (a) purification using the QIAquick Nucleotide Removal Kit are shown.|GelPilot Loading Dye contains three tracking dyes to facilitate optimization of DNA resolution.
|QIAcube.
|Microcentrifuge.
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Performance
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.
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 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").

Procedure

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, enabling increased productivity and standardization of results (see figures "Spin column handling options A, and B").

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, and microinjection.

Feature
Specifications
Binding capacity 10 µg
Elution volume 30–50 µl
Format Tube
Fragment size 40 bp – 10 kb
Processing Manual
Recovery: oligonucleotides dsDNA Recovery: oligonucleotides, dsDNA
Removal <10mers 17–40mers dye terminator proteins Removal <10mers
Sample type: applications DNA, oligonucleotides: PCR reactions
Technology Silica technology

Supplementary Protocols
1
QIAquick® Spin Columns can now be used on any vacuum manifold with luer connectors, for example, the QIAvac 6S or QIAvac 24 with QIAvac Luer Adapters. This protocol is designed for removal of primers <10 bases, enzymes, salts, and unincorporated nucleotides from biotin-, or DIG-labeled DNA fragments and oligonucleotides >17 nucleotides.
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