Member since: 2019
Organization: University of Liverpool
Good detailed information online for troubleshooting
Application Area: DNA extraction
"Extremely easy to use (beginner/ undergrad/research level) with simple detailed instructions and high success rates. Good detailed information online for troubleshooting. Multi-use product and relatively quick. Only downside is the expense at £200. "
The PureLink® HiPure Plasmid Filter Maxiprep Kit is designed to isolate plasmid DNA of the highest purity from 100 to 500 mL of an overnight bacterial culture. The PureLink® HiPure Plasmid Filter Kits include HiPure Filter Columns, which combine filtration for clarification of the bacterial lysate with anion-exchange resin purification of plasmid DNA, in a single unit. The resulting plasmid DNA is of a quality equivalent to that obtained by purification with two passes through cesium chloride gradients—the most rigorous method for DNA purification. Purify high-quality DNA in less than two hours for transfections and other applications with no need for additional steps to remove contaminants like RNA, proteins, or endotoxins. Additionally, the elimination of phenol, chloroform, ethidium bromide, and cesium chloride from the protocol minimizes exposure to, and disposal of hazardous materials. The PureLink® HiPure Plasmid Filter Kits provide:
• Simple protocols—bacterial lysate clarification without centrifugation
• Fast results—save time by purifying plasmid DNA in less than two hours
• High yields—up to 200 µg for midipreps and up to 850 µg for maxipreps (see table)
• Reliable performance—the high-quality plasmid DNA purified with these kits is suitable for a wide variety of applications
How it works
The addition of a HiPure Filter Column to the PureLink® HiPure Plasmid Kits provides rapid clearing of the bacterial lysate without the need for centrifugation (see figure). The lysate filtration cartridge is integrated into the DNA-binding column for a combined step of lysate clarification and plasmid DNA binding directly to the anion-exchange resin. A single column wash removes impurities such as RNA, proteins, metabolites, and other low molecular weight molecules; the ultrapure plasmid DNA is then eluted with a high-salt buffer. The DNA is then desalted and concentrated with an isopropanol precipitation step, then collected by centrifugation. The entire protocol can be completed in about two hours.