The GenElute HP Endotoxin-Free Plasmid Maxiprep Kit offers a simple and rapid method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. The kit uses a vacuum format with a filter column for the rapid clearing of the bacterial lysate and a silica column for capturing plasmid DNA. A proprietary solution binds plasmid DNA to the binding column while preventing endotoxins from adsorbing. The technology allows for the user to consistently achieve levels of endotoxin that are less than 0.1 endotoxin units per μg of plasmid DNA.
High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells.
GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit has been used in the isolation of plasmid from swine liver and from competent Escherichia coli cells.
Features and Benefits
- Up to 1.2 mg of high-copy plasmid DNA with endotoxin levels of ≤0.1 EU/μg
- Only 35 minutes from pelleted cells to purified plasmid
- Convenient vacuum format
An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is clarified by filtration followed by the addition of a binding solution that has been optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured on silica, while endotoxins are prevented from adsorbing to the membrane. Two wash steps remove contaminants. Finally, the bound DNA is eluted in endotoxin-free water. The recovered plasmid DNA is predominately in its supercoiled form. Genomic DNA and RNA are below detectable levels by ethidium bromide stained agarose gel electrophoresis.
GenElute is a trademark of Sigma-Aldrich Co. LLC