QIAquick 96 PCR Purification Kits
For purification of 96 PCR products (up to 10 μg), 100 bp to 10 kb
- Up to 95% recovery of ready-to-use DNA
- Fast and convenient procedure
- Cleanup of DNA up to 10 kb in three easy steps
Cat. No. / ID: 28181
QIAquick 96 PCR Purification Kits provide 96-well plates, buffers, and collection tubes for high-throughput silica-membrane-based purification of PCR products >100 bp in size. DNA up to 10 kb is purified using a simple and fast bind–wash–elute procedure and an elution volume of 60–80 μl (resulting in an eluate volume of 40–60 μl). The cleanup procedure can be fully automated on the BioRobot workstations using the QIAquick 96 PCR BioRobot Kit.
QIAquick 96 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. 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.
The QIAquick 96 procedure allows parallel purification of up to 96 PCR samples using efficient vacuum-driven purification on a QIAvac 96.
The QIAquick 96 PCR BioRobot Kit is a special kit format optimized for use on the BioRobot 9600 (no longer available), the BioRobot 3000 (no longer available) and the BioRobot 8000. The kit provides QIAquick 96 modules, together with all buffers and plasticware required for automated high-throughput cleanup of 96 PCR samples.
The QIAquick system uses a simple bind–wash–elute procedure (see flowchart " QIAquick 96 procedure"). Binding buffer is added directly to the PCR sample or other enzymatic reaction, and the mixture is applied to the 96-well plate. 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 subsequent applications.
QIAquick multiwell modules are processed using vacuum-driven purification on QIAvac manifolds. The QIAquick 96 PCR Purification Kit requires the use of the QIAvac 96 vacuum manifold. The cleanup procedure can be fully automated on the BioRobot workstations using the QIAquick 96 PCR BioRobot Kit.
DNA fragments purified with the MinElute or QIAquick system are ready for direct use in many applications, including sequencing, microarray analysis, ligation and transformation, restriction digestion, labeling, microinjection, PCR, and in vitro transcription.
|Binding capacity||10 µg|
|Removal <10mers 17–40mers dye terminator proteins||Removal <40mers|
|Sample type: applications||DNA, oligonucleotides: PCR reactions|
|Fragment size||100 bp – 10 kb|
|Recovery: oligonucleotides dsDNA||Recovery: oligonucleotides, dsDNA|
|Elution volume||60–80 µl|
Larger DNA fragments bind more tightly to the QIAquick columns. It is difficult to predict whether a DNA fragment larger than 10 kb can be efficiently recovered, because this depends on base composition as well as fragment size. If the fragment is only a few kb larger than the 10 kb limit, it can be helpful to heat the elution buffer EB to 60°C and let it incubate on the column for a few minutes before centrifuging. However, please note that it will become less likely to recover your sample the larger the fragment size is. As we cannot guarantee recovery of fragments larger than the maximum cutoff size, we do not recommend to purify such fragments using QIAquick Kits.
The QIAEX II Kit can be used to extract DNA fragments up to 50 kb from agarose or polyacrylamide gels.
It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.
Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.
Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.
Buffer PE did not contain ethanol
Ethanol must be added to Buffer PE (concentrate) before use. Repeat procedure with correctly prepared Buffer PE.
Inappropriate elution buffer
DNA will only be eluted efficiently in the presence of low-salt buffer (e.g., Buffer EB: 10 mM Tris·Cl, pH 8.5) or water. Elution efficiency is strongly dependent on the salt concentration and pH of the elution buffer. Contrary to adsorption, elution is most efficient under basic conditions and low salt concentrations. DNA is eluted with 50 or 30 µl of the provided Buffer EB (10 mM Tris·Cl, pH 8.5), or water. The maximum elution efficiency is achieved between pH 7.0 and 8.5. When using water to elute, make sure that the pH is within this range. In addition, DNA must be stored at –20°C when eluted with water since DNA may degrade in the absence of a buffering agent. Elution with TE (10 mM Tris·Cl, 1 mM EDTA, pH 8.0) is possible, but not recommended because EDTA may inhibit subsequent enzymatic reactions.
Elution buffer incorrectly dispensed
Add elution buffer to the center of the QIAquick membrane to ensure that the buffer completely covers the membrane. This is particularly important when using small elution volumes (30 µl).
Yes, please follow the Supplementary Protocol 'Spin procedure for purifying the 2x96 PCR samples using the Plate Rotor 2x96, a special centrifuge and the QIAquick 96 PCR Purification Kit' (QQ01). Please contact your local QIAGEN Technical Service for this protocol.
Yes, please follow the Supplementary Protocols 'High-throughput gel extractions using the QIAquick 96 PCR Purification Kit' (QQ03). Please contact your local QIAGEN Technical Service for this protocol.
DNA fragments purified with the QIAGEN DNA Cleanup Systems, i.e., the QIAquick PCR Purification Kit, the MinElute Reaction Cleanup Kit, the QIAEX II Gel Extraction Kit etc. may float out of the loading wells of agarose gels due to residual ethanol carried over from the wash step with Buffer PE (despite the addtition of glycerol-containing loading buffer).
Use either of the following options to remove residual ethanol from the eluate: