Other Notes
Endotoxins (also known as lipopolysaccharides or LPS) are often co-purified with plasmid DNA and can significantly reduce transfection efficiencies in sensitive cell lines. The GenElute Endotoxin-free Plasmid Kits offer a simple, rapid, cost-effective method for purifying plasmid DNA with ≤0.1 EU/μg DNA for high efficiency transfection.
Features and Benefits
• Higher transfection efficiency and yield than market leaders
• Purify up to 1.2 mg endotoxin-free plasmid DNA
(≤0.1 EU/ug DNA)
• Fast and simple allowing up to 12 preps in less than 2 hours
• Faster than gravity flow ion exchange columns or magnetic bead preps
• No expensive magnetic equipment required
Principle
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 with the Midiprep Kit (Product Code PLED35) and up to 1.2 mg from 25 to 130 mL of culture with the Maxiprep Kit (Product Code PLEX15).
The recovered plasmid DNA is predominately in its supercoiled form. There is no visual evidence of genomic DNA or RNA contamination on ethidium bromide-stained agarose gels. The DNA is ready for immediate use in downstream applications such as transfection, restriction endonuclease digestion, cloning, sequencing, and PCR amplification.
Legal Information
GenElute is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.
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