The PureLink® HiPure Plasmid Purification Systems are designed to isolate transfection-grade plasmid DNA from E. coli. The Midiprep Kit protocol will typically yield 100–350 µg plasmid DNA from 15–25 ml bacterial culture, at a purity that is comparable to that achieved by two passes through cesium chloride gradients.
Advantages of the PureLink® HiPure Plasmid Midiprep Kit include:
• Labor- and time-saving—No need for extra steps to remove endotoxin and other contaminants
• High purity, low endotoxin product—pure enough for mammalian cell transfection
• Versatility—purify any type and size of plasmid DNA, including BAC, bacmids, and ssM13 DNAs
Anion-exchange chromatography for plasmid purification
The PureLink™ HiPure Plasmid DNA Purification Kits use a patented anion-exchange resin to purify plasmid DNA to a level equivalent to two passes through CsCl gradients. The resin combines excellent capacity with a fast flow rate, high resolution, high yield, and efficient endotoxin removal. Plasmid preparations can typically be completed in less than 2 hours.
Unlike cesium chloride gradient protocols, PureLink® HiPure Plasmid Purification Systems do not employ organic solvents, ethidium bromide, or cesium chloride, all of which are troublesome to work with and dispose. The DNA purity that you expect from a CsCl gradient, however, is still delivered. Typically, plasmid DNA prepared using the PureLink® HiPure Plasmid Midiprep Kit will have a A260/A280 ratio of >1.80, indicating that the DNA is reasonably free of proteins that might interfere with downstream applications. Endotoxin levels are typically within 0.1–1 EU/ µg, making this plasmid DNA ideal for mammalian cell transfection.