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.
We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product has Inherently Safer Chemistry, compared to the standard use of phenol and chloroform to perform DNA extractions.