Merck, PLED35-1KT, GenElute™ Endotoxin-free Plasmid Midiprep Kit

The GenElute Endotoxin-free Plasmid Kit offers a simple, rapid, cost effective method for purifying plasmid DNA with ≤0.1 EU/μg DNA for high efficiency transfection. Endotoxins (also known as lipopolysaccharides or LPS) are often co-purified with plasmid DNA and significantly reduce transfection efficiencies in endotoxin-sensitive cell lines. Recombinant E. coli is harvested from an overnight culture by centrifugation and subjected to a modified alkaline-SDS lysis procedure to produce a cleared lysate. Endotoxins are removed from the cleared lysate with simple extraction-phase separation steps. Plasmid DNA is further purified by adsorption onto silica in the presence of high salts. After a simple spin-wash step, the bound plasmid DNA is eluted in endotoxin-free water. Up to 250 μg of endotoxin-free plasmid DNA may be prepared from 5 to 40 mL of culture. The recovered plasmid DNA is predominately in its supercoiled form. The DNA is ready for immediate use in downstream applications such as transfection, restriction digestion, cloning, sequencing, and PCR amplification.