The NucleoSpin® 8/96 Plasmid procedure is a modified version of the Birnboim and Doly alkaline lysis plasmid miniprep protocol. Bacterial cultures are harvested by an initial centrifugation step. After resuspension of the pelleted bacteria (Buffer A1) and alkaline cell lysis (Buffer A2), a neutralization and binding buffer (Buffer A3) containing chaotropic salts is added. Resulting bacterial crude lysates are cleared by vacuum filtration with the NucleoSpin® Plasmid Filter Strips/Plate. The cleared lysates containing the plasmid DNA are collected into the NucleoSpin® Plasmid Binding Strips/Plate. The chaotropic salt leads to a reversible adsorption of the plasmid DNA to the NucleoSpin® silica membrane during the second vacuum-filtration step. High purity of the final plasmid DNA preparation is achieved by complete removal of cellular contaminants, salts, detergents, and other compounds by subsequent washing steps. Highly pure plasmid DNA is finally eluted with Elution Buffer AE (5 mM Tris / HCl, pH 8.5) or water (pH 8.0 – 8.5), and can directly be used for downstream applications.

This kit is intended for use under vacuum. A support protocol for elution under centrifugation is included in the manual. A support protocol for complete processing under centrifugation is available from our Technical Service.