4/7/2023 0 Comments Quick quotes page kitsTo help dissolve the gel, mix by vortexing the tube every 2–3 min during the incubation. Add 6 volumes of Buffer QG to 1 volume of gel, based on the gel weight (100 mg ~ 100 µl). Remove the gel slice from the TE buffer, and place it in a colorless tube.Soak the gel slice in TE buffer for 25 min at room temperature with gentle shaking. Weigh the gel slice, and record the weight.Excise the RNA fragment from the formaldehyde agarose gel with a clean, sharp scalpel.QIAquick Gel Extraction Kits are not guaranteed to be RNase-free. Note that this protocol has not been thoroughly tested and optimized by QIAGEN. Knobloch, Heinrich Heine University, Düsseldorf, Germany. The QIAquick Gel Extraction Kit for extraction of DNA from gels can also be used for RNA gel extraction. Please see a user-developed procedure below, which was kindly provided by J. This is particularly important when using small elution volumes (30 µl). Elution with TE (10 mM Tris♼l, 1 mM EDTA, pH 8.0) is possible, but not recommended because EDTA may inhibit subsequent enzymatic reactions.Īdd elution buffer to the center of the QIAquick membrane to ensure that the buffer completely covers the membrane. In addition, DNA must be stored at –20☌ when eluted with water since DNA may degrade in the absence of a buffering agent. When using water to elute, make sure that the pH is within this range. The maximum elution efficiency is achieved between pH 7.0 and 8.5. DNA is eluted with 50 or 30 µl of the provided Buffer EB (10 mM Tris♼l, pH 8.5), or water. Contrary to adsorption, elution is most efficient under basic conditions and low salt concentrations. Elution efficiency is strongly dependent on the salt concentration and pH of the elution buffer. Repeat procedure with correctly prepared Buffer PE.ĭNA will only be eluted efficiently in the presence of low-salt buffer (e.g., Buffer EB: 10 mM Tris♼l, pH 8.5) or water. See figuresĮthanol must be added to Buffer PE (concentrate) before use. The QIAquick Gel Extraction 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, B, C, D, and E"). The spin columns fit into a conventional table-top microcentrifuge or onto any vacuum manifold with luer connectors, such as the QIAvac 24 Plus with QIAvac Luer Adapters. QIAquick spin columns are designed to provide two convenient handling options. 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. Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the QIAquick spin column (see figure " pH Indicator Dye"). The QIAquick system uses a simple bind-wash-elute procedure (see flowchart " QIAquick and MinElute procedure").
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |