Ion exchange purification of 45-100 µg plasmid DNA from 15-100 ml of E. coli cells.
PhoenIX™ plasmid purification systems from Qbiogene rely on ion-exchange chromatography through a unique, patented solid-phase column. Precisely balanced pH values and salt concentrations permit optimal purification of highly clean plasmid DNA from the unique ion-exchange resin. Unlike resins found in other ion-exchange kits, PhoenIX columns are distinct: First, the smaller pore size of the resin particles increases the surface area available for binding. Second, the spacer molecule linking the positively charged amino group to the resin particles is longer than in other resins, allowing better separation between different biomolecules during the binding and washing process.
The PhoenIX Midiprep Kits and PhoenIX Maxiprep Kits rely on centrifugation to clear the bacterial lysate, and on gravity to pull liquids through the column material. The PhoenIX Gigaprep Kit is driven by vacuum and offers lysate clearing via a filtration cartridge.
Plasmid DNA purified with PhoenIX columns can be used immediately in a wide variety of downstream applications including automated fluorescent sequencing, enzymatic amplification, labeling, transcription, cloning and other enzymatic manipulations. Additionally, the levels of endotoxin that remain after purification are so low that successful transfection will occur with even sensitive cell lines.
Two protocols are included in this instruction manual. The "high copy" protocol allows isolation of plasmid DNA from bacterial cultures of 15 ml to 50 ml. The "low copy" protocol allows isolation of plasmid DNA from bacterial cultures of 51 ml to 100 ml. In either protocol, plasmid DNA is ready to precipitate in just over an hour.
- Patented ion exchange resin and optimized buffer system offers:
- Increased binding capacity for higher yield
- Superior separation of unwanted molecules during wash step
- Elution of ultrapure plasmid equal to 2 passes through CsCl
- Consistent, reproducible results from column to column
- Fast protocol that saves at least an hour over similar kits
- Purification of constructs up to 150 kb
- Safe procedure with no hazardous materials
- Low endotoxin levels allow transfection without additional purification
- Pre-packed columns flow at a consistent, even rate with no clogging
- Column support rack included in kit eliminates need for seperate purchase
- Low price and flexibility add value to lab budgets and procedures
Each of the 25 preps in the PhoenIX Midiprep Kit provides 100 µg of ultrapure plasmid DNA from 15 - 100 ml of bacterial culture. Cleared bacterial lysate is prepared by centrifugation and loaded onto the PhoenIX Midi Column for rapid purification by gravity flow. Protocols for high copy and low copy plasmids are included in the instruction manual.
Kit Components: RNase A, Cell Resuspension Buffer, Lysis Buffer, Neutralization Buffer, PhoenIX Midi Columns, Disposable Midi Column Rack, Equilibration Buffer, Column Wash Buffer, Elution Buffer.