The starting material is lysed by incubation in a solution containing SDS and Proteinase K. Once lysis is complete, chaotropic salts are then added to further promote solubilization of macromolecules. The next step is to add ethanol which ensures that the appropriate binding conditions are created to bind the DNA to the silica membrane. Once the DNA is bound to the silica membrane several wash buffers are used to remove contaminants. The final step is to elute pure genomic DNA in the Tissue Elution Buffer included in the kit.
Store the components of the GenElute™ 96 Well Total DNA Purification Kit at room temperature. If any regent forms a precipitate, warm at 30-40 °C until the precipitate dissolves and allow to cool to room temperature before use. All reagents are stable for at least one year when stored properly.
Before beginning the procedure, complete the following:
Establish a reliable vacuum source for the GenElute™ Vacuum Manifold. The manifold may be used with a vacuum pump, house vacuum, or water aspirator. A vacuum of 200-400 mbar (pressure difference) is recommended. The use of a GenElute™ Vacuum Regulator (VM02) is also recommended. Alternatively, adjust vacuum during the purification such that the sample flows through the column with a rate of 1-2 drops per second. To check the vacuum in advance it is recommended to place a Tissue Binding Plate on top of the vacuum lid. Seal the Binding Plate with common self-adhering foil. Close the manifold’s valve and check and adjust the vacuum pressure.
This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices. Proteinase K is an irritant to eyes, respiratory system, and skin. It may also cause sensitization by inhalation. Gloves and safety glasses should be worn. Tissue Binding Buffer and Tissue Wash Buffer 1 contain guanidinium hydrochloride which is an irritant so gloves and safety glasses should be worn.