PureLink™ HiPure Plasmid FP (Filter and Precipitator) Maxiprep Kit
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The PureLink™ HiPure Plasmid FP (Filter and Precipitator) Maxiprep Kit isolates plasmid DNA, from cell pellet to ultrapure DNA, in less than one hour. Using the HiPure Filter Columns and HiPure Precipitators supplied in the kit, you can purify plasmid DNA without the centrifugation steps required in traditional isolation protocols. The quality of the purified plasmid DNA is equivalent to that obtained by two passes through cesium chloride gradients - the most rigorous method for DNA purification - making it suitable for transfection and other applications requiring high-purity plasmid DNA. The PureLink™ HiPure Plasmid FP Maxiprep Kit (Table 1) provides: • Simple protocols - bacterial lysate clarification and isopropanol precipitation without centrifugation • Fast results - from E. coli cell pellet to high-purity, transfection-grade plasmid DNA in about 60 minutes • High yields - up to 850 µg from a high-copy plasmid
How it works The PureLink™ HiPure Plasmid FP Maxiprep Kit employs an alkaline lysis method to prepare and rapidly clear bacterial lysates without the need for centrifugation (Figure 1). The unique HiPure Filter Column includes a lysate filtration cartridge integrated into the anion-exchange column for single-step lysate clarification and plasmid DNA binding. Impurities such as RNA, proteins, and other contaminants are removed with a single wash, and ultrapure plasmid DNA is eluted with a high-salt buffer. To precipitate and desalt the DNA, isopropanol is added to the eluted DNA and the mixture is then applied to the HiPure Precipitator using a large syringe. After a subsequent washing and drying step, the plasmid DNA is easily eluted from the HiPure Precipitator with either TE buffer or water. The use of the precipitator eliminates the need for centrifugation, reducing the risk of losing the DNA pellet during supernatant removal, and saving time. The resulting DNA is ready to use in the most challenging applications. The entire protocol - from E. coli cell pellet to ultrapure transfection-grade plasmid DNA - can be completed in about an hour.
Highest yields and ready-to-use concentrations The HiPure Precipitator has been optimized to allow for lower elution volumes (down to 500 µl) to produce highly concentrated DNA without a significant decrease in yields. In fact, yields at higher concentrations are consistently higher than the competitor kits (Figure 2). Yields and concentration can be adjusted to meet the needs of your downstream application.
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