QIAquick 96 PCR Purification Kits

For purification of 96 PCR products (up to 10 µg), 100 bp to 10 kb

S_1346_DNA_QQ0827

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QIAquick 96 PCR Purification Kit (4)

Cat. No. / ID:   28181

For purification of 4 x 96 PCR reactions: 4 QIAquick 96 Plates, Buffers, Collection Microtubes (1.2 ml), Caps
Kit
QIAquick 96 PCR Purification Kit
QIAquick 96 PCR BioRobot Kit
Preparations
4
24
QIAquick 96 PCR Purification Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Up to 95% recovery of ready-to-use DNA
  • Fast and convenient procedure
  • Cleanup of DNA up to 10 kb in three easy steps

Product Details

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 Universal workstation using the QIAquick 96 PCR BioRobot Kit.

Performance

The QIAquick 96 PCR Kit provides a quick and easy method for high-throughput purification of PCR samples, with a recovery rate of up to 90%. The QIAquick 96 procedure delivers highly pure DNA suitable for various downstream applications (see figure " Accurate sequencing"). Using the QIAvac 96, DNA fragments 100 bp – 10 kb in size are purified from 1–4 x 96 samples.
See figures

Principle

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 Universal. The kit provides QIAquick 96 modules, together with all buffers and plasticware required for automated high-throughput cleanup of 96 PCR samples.

Procedure

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.

Handling

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 Uworkstations using the QIAquick 96 PCR BioRobot Kit.

See figures

Applications

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.

Supporting data and figures

Specifications

FeaturesSpecifications
Binding capacity10 µg
ProcessingManual/automated
Removal <10mers 17–40mers dye terminator proteinsRemoval <40mers
Sample type: applicationsDNA, oligonucleotides: PCR reactions
Format96-well plate
Fragment size100 bp – 10 kb
TechnologySilica technology
Recovery: oligonucleotides dsDNARecovery: oligonucleotides, dsDNA
Elution volume60–80 µl

Resources

Quick-Start Protocols (1)
Technical Information and Important Notes (2)
Kit Handbooks (1)
For rapid purification of multiple PCR products 
Safety Data Sheets (1)
Certificates of Analysis (1)

Publications

Temporal and differential gene expression of Singapore grouper iridovirus.
Chen LM; Wang F; Song W; Hew CL;
J Gen Virol; 2006; 87 (Pt 10):2907-2915 2006 Oct PMID:16963749
Comparative genomics of host-specific virulence in Pseudomonas syringae.
Sarkar SF; Gordon JS; Martin GB; Guttman DS;
Genetics; 2006; 174 (2):1041-56 2006 Sep 1 PMID:16951068
cDNA microarrays as a tool for identification of biomineralization proteins in the coccolithophorid Emiliania huxleyi (Haptophyta).
Quinn P; Bowers RM; Zhang X; Wahlund TM; Fanelli MA; Olszova D; Read BA;
Appl Environ Microbiol; 2006; 72 (8):5512-26 2006 Aug PMID:16885305
Characterization of the vernalization response in Lolium perenne by a cDNA microarray approach.
Ciannamea S; Busscher-Lange J; de Folter S; Angenent GC; Immink RG;
Plant Cell Physiol; 2006; 47 (4):481-92 2006 Jan 31 PMID:16449231
Anti-inflammatory activity in vitro and in vivo of the protein farnesyltransferase inhibitor tipifarnib.
Xue X; Lai KT; Huang JF; Gu Y; Karlsson L; Fourie A;
J Pharmacol Exp Ther; 2005; 317 (1):53-60 2005 Dec 13 PMID:16352705

FAQ

I bound an 11 kb DNA fragment to a QIAquick column; is it completely lost?

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.

 

FAQ ID -756
How can I improve recoveries when using the QIAquick Kits?

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).

FAQ ID -180
Can I use a centrifuge instead of vacuum when using the QIAquick 96 PCR Purification Kit?
Yes, you can use a QIAGEN Centrifuge with the QIAquick 96 PCR Purification Kit. Follow the Supplementary Protocol 'Spin procedure for purifying 2 x 96 PCR samples using the Plate Rotor 2 x 96, a special centrifuge, and the QIAquick 96 PCR Purification Kit.' Here's the link to the protocol.
FAQ ID -293
Do you have a protocol for purifying 2x96 PCR samples simultaneously with the QIAquick 96 PCR Purification Kit?

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.

The protocol is for use with QIAGEN Centrifuges and the Plate Rotor 2 x 96.

FAQ ID -935
Do I have to remove the oil from my PCR reaction before using the QIAquick or MinElute PCR Purification Kit?
No - mineral oil will not affect the clean-up procedure with the QIAquick or MinElute PCR Purification Kit.
FAQ ID -575
Do you have protocols for multiple extractions of DNA fragments from agarose gels?

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.

FAQ ID -944
Why does my DNA sample float out of the slot when loading it onto an agarose gel?

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:

  • re-purify the sample using a QIAquick-, or MinElute column, or QIAEX II resin
  • incubate the eluate at 56°C for 10 min to evaporate the ethanol
  • dry down the sample in a vacuum centrifuge, and resuspend the pellet in a small volume of sterile water
FAQ ID -205