An overnight recombinant E. coli culture is harvested with centrifugation and subjected to a modified alkaline-SDS lysis procedure. Bacterial cells are harvested by centrifugation and resuspended in Resuspension Solution containing RNase. The cells are then lysed and neutralized followed by the addition of Binding Solution. The prepared lysate is then transferred to a Sigma HP Filter Plate assembled in a vacuum manifold where it is filtered and collected into a deep well plate. The vacuum manifold is rearranged and the filtered lysate is transferred to the Sigma HP Binding Plate. The plasmid DNA is captured onto the silica based membrane and impurities are removed by successive wash steps. Finally, the binding plate is dried and the plasmid DNA is eluted in Elution Solution.
The GenElute HP 96 Well Plasmid Miniprep Kit offers a simple, rapid, costeffective solution for high-throughput purification of plasmid DNA from recombinant E. coli cultures. By combining silica-binding technology and the convenience of a vacuum format, up to 10 μg of high copy plasmid DNA per well can be recovered from 1.3 mL of culture in less than 50 minutes. Actual yields and the optimum volume of culture depend on the plasmid and culture medium used.