Plasmid DNA Purification—Automated Processing 96-Well Plates—NucleoSpin Robot-96 Plasmid
The NucleoSpin Robot 96 Plasmid Kit allows fully automated purification of high-copy plasmids from E. coli, with common liquid handling instruments. Manual interactions are not required. Ready-to-run protocols are available at MACHEREY-NAGEL and can be customized to fit your needs. The robot kits allow the rapid, small-scale purification of up to 20 µg of plasmid DNA per preparation from 1–5 ml of E. coli cultures grown overnight in square-well plates or tubes. E. coli cultures can be grown in a volume of up to 1.5 ml using the supplied culture plate. E. coli cultures grown in up to 5 ml LB, 3 ml 2YT, or 3 ml TB can be processed when using suitable culture tubes or plates (not supplied with the kit).
The NucleoSpin Robot 96 Plasmid Kit uses advanced silica-membrane technology in combination with efficient lysate filtration. Bacterial cultures are pelleted by centrifugation and cells are lysed under alkaline conditions. A buffer containing large amounts of chaotropic ions is added to neutralize the lysate and to allow reversible adsorption of DNA to the NucleoSpin silica membrane. Crude lysates are cleared with the NucleoSpin Plasmid Filter Plates, and the DNA is reversibly bound to the silica membrane in the NucleoSpin Plasmid Binding Plates. Contaminants are removed by subsequent washing steps. An additional washing with high-salt buffer AW is recommended for E. coli host strains with high endogenous nuclease activity, for long range DNA sequencing, and for better performance of critical enzymatic reactions. Highly pure plasmid DNA is eluted with low-salt elution buffer (5 mM Tris/HCl pH 8.5) or water (pH 8.0–8.5), and is ready-to-use for further applications.
NucleoSpin 96 Plasmid Core Kits: Reduced kit composition plus a large variety of accessories (available separately) allow for optimal adjustment of the kits to individual needs.