Geneaid Midi Plasmid Kit (Endotoxin Free)

The Geneaid™ Midi Plasmid Kit Endotoxin Free uses pre-packed anion-exchange resin columns to purify plasmid DNA from 50-300 ml of cultured bacterial cells. TrueBlue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. Using an efficient gravity-flow procedure, plasmid DNA is bound to the column and contaminants are efficiently removed. The purified plasmid DNA is eluted then precipitated with isopropanol for desalting. The entire procedure can be completed without ultracentrifuges, HPLC or other toxic reagents and the purified plasmid DNA is suitable for transfection, sequencing reactions, ligation, PCR, in-vitro transcription, microinjection, restriction enzyme digestion and gene gun.

Advantages (Cat. # PIE25)

• High Yield: >500 µg of pure transfection grade plasmid DNA from 100 ml of cultured bacterial cells
• Efficient endotoxin removal (<0.1 EU/µg DNA) verified by LAL
• Sample Volume: 50-300 ml of cultured bacterial cells
• Format: plasmid midi anion-exchange resin column
• Operation time: within 2 hours (only 80 minutes when combined with the Presto™ Plasmid Concentrator)
• Storage: dry at room temperature (15-25ºC) for up to 2 years

Applications

Transfection, Sequencing, Ligation, PCR, In-vitro Transcription, Microinjection, Restriction Enzyme Digestion, Gene Gun

Components

• PM1, PM2, PM3 Buffer
• TrueBlue Lysis Buffer
• PER Buffer
• PEQ Buffer
• PW Buffer
• PEL Buffer
• RNase A
• Plasmid Midi Columns

Quality Control

The quality of the Geneaid™ Midi Plasmid Kit (Endotoxin Free) is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Plasmid DNA from a 50 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the Geneaid™ Midi Plasmid Kit. Following the purification process, a yield of more than 300 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid DNA (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.