High Purity Plasmid Purification Kits

Marligen's High Purity Plasmid Purification Kits are designed for preparing ultrapure plasmid DNA equivalent to the gold standard of two cycles of CsCl gradient purification and are suitable for all applications. Mini, midi and maxiprep columns employ gravity flow, while mega and gigaprep formats use vacuum-assisted gravity flow. Marligen now offers midi and maxiprep kits with a convenient integrated prefilter in the columns for even greater convenience and speed. Neutralized lysates can be added directly to columns containing the prefilters, thereby combining lysate clarification and column loading into one simple step.

The procedure employs modified alkaline lysis and neutralization followed by ion exchange chromatography using a patented resin designed to provide excellent capacity, resolution and endotoxin removal (Fig. 1). DNA purified with Marligen's High Purity Kits is suitable for all applications. Purity, integrity and performance in restriction endonuclease digestion and ligation are verified by agarose gel electrophoresis (Fig. 2). Figure 3 demonstrates that high quality DNA prepared with these kits performs exceptionally well in transfection experiments, even with difficult cell lines such as Huh7.

Fig. 1 The Marligen High Purity resin is composed of smaller particles with more uniform pore size than competitors' products, providing greater yield and reproducibility. A longer spacer arm provides greater DNA binding efficiency and the unique patented ion-exchange moiety increases the efficiency of separation of RNA and DNA	Fig. 2 Plasmid DNA (pBluescript IIKS) was prepared using the Marligen High Purity Plasmid Purification Kits. The DNA was digested with EcoR1 and ligated with a 1.65 kb DNA fragment. Lane 1: 1 kb Ladder Lane 2: DNA before ligation Lane 3-5: Ligation with 0.02, 0.05 and 0.01 units of Eco R1.	Fig. 3 This comparison of transfection efficiency with several cell lines using DNA prepared by various methods demonstrates that Marligen's Prep yields DNA that offers superior performance in transfection.