QIAquick 96 PCR Purification Kits

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.

 

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, 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.

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.

No - mineral oil will not affect the clean-up procedure with the QIAquick or MinElute PCR Purification Kit.