Midi preparation of plasmid DNA with transfection-grade purity using vacuum

Endotoxins are lipopolysaccharides included in the cell wall of gram-negative bacteria. Endotoxins are co-purified during standard plasmid preparations resulting in lower survival rate of transfected eukaryotic cells. MACHEREY-NAGEL has developed a new technology to reduce endotoxins from bacterial lysates in a convenient silica membrane based preparation. The NucleoSnap Plasmid Midi kit is based on alkaline lysis to liberate plasmid DNA from the bacterial cells. The kit includes a NucleoSpin® Plasmid Filter Column for lysate clarification by centrifugation. Endotoxins and proteins are removed during the wash step by the innovative Detoxification Buffer. Additionally, the kit is based on a new innovative column design composed of an upper part that can hold up to 30 mL E.coli lysate and a bottom part which is a mini spin column for DNA concentration. After loading the cleared lysate and the wash steps with the large volumes on a vacuum manifold, the upper part can ...