Details
The Maxi Plasmid Kits use pre-packed ion-exchange resin columns to purify plasmid or cosmid DNA from bacterial cultures. In this process, the modified alkaline lysis method and RNase treatment are used for creating cleared lysate with minimal genomic DNA and RNA contaminants. Using a gravity-flow procedure, the plasmid DNA in crude lysate has been bound to the column. The contaminants can be washed off with a wash buffer. Finally, the purified DNA is eluted by a high salt buffer and then precipitated with isopropanol for desalting. The entire procedure can be completed in less than two hours. The purified plasmid DNA is suitable for transfection, sequencing reactions, PCR, and in-vitro transcription.
The quality of the Fast-Ion Maxi Kits is tested on a lot-to-lot basis by isolating plasmid DNA from a 100 ml overnight E. coli (DH5α) culture, containing pBluescript (A600>2U/ml). More than 400 µg of plasmid DNA is quantified with a spectrophotometer. The purified plasmid (1 µg) is used in EcoRI digestion, and checked by electrophoresis.
Features:
• Sample Size: 100 mL high-copy plasmid/250 mL low-copy plasmid
• Operation: Gravity Flow Column
• Expectant Yield: 500 µg to 1 mg of plasmid DNA
• Operation Time: 120 minutes or less