Norgen’s EndotoxinRemoval Kit is designed for the rapid removal of endotoxins from up to 1 mg of previously purified DNA. Endotoxins, also known as lipopolysaccharides, are cell-membrane components of Gram-negative bacteria such as E. coli. Endotoxins are released during the lysis step of plasmid purification and significantly reduce transfection efficiencies in endotoxin sensitive cell lines. Therefore, the removal of endotoxins from plasmid preparations is often necessary prior to the use of the DNA in downstream applications. With Norgen’s Endotoxin Removal Kit, endotoxin levels are efficiently reduced to 0.1 EU/μg DNA or less. Each kit contains sufficient materials for 4 purifications, and preparation time for a single sample is approximately 30 minutes.
Norgen’s Purification TechnologyPurification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The plasmid DNA is preferentially purified from the contaminating endotoxins with this kit. The first step in the process involves the addition of Binding Solution to the DNA sample (please see flow chart on page 3). The sample is then placed into the top reservoir of the column, and a small amount of Endotoxin Removal Solution is added. After a brief incubation, isopropanol is also added to the column and the solution is mixed and then spun in a centrifuge. Norgen’s resin binds DNA in a manner that depends on ionic concentrations, thus only the plasmid DNA will bind to the column while the contaminating endotoxins will be removed in the flowthough. The bound DNA is then washed once with the provided wash buffer to remove any remaining impurities. Lastly, the endotoxin-free plasmid DNA is eluted with the elution buffer. The purified DNA is of the highest quality and can be used in a number of downstream applications including sequencing, cloning, and transfections.