I-Blue Midi Plasmid Kits optimize isolation of plasmid DNA from E.coli with a simple lysis, bind, wash, and elute protocol. The kits include I-Blue lysis buffer, which helps prevent common handling errors, ensuring efficient cell lysis and SDS precipitation.
- Sample Size: 50 – 150 ml of cultured bacterial cells
- Operation: anion-exchange resin column, gravity flow
- Expectant Yield: up to 400 µg of pure transfection grade plasmid DNA form 100 ml cultured bacterial cells
- Elution Volume: 500 µl – 2ml
- Operation Time: 80 minutes or less
The I-Blue Midi Plasmid Kit uses pre-packed anion-exchange resin columns to purify plasmid DNA from 50 – 150 ml of cultured bacterial cells. I-Blue Lysis Buffer (an option 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. I-Blue lysis buffer is added during the cell re-suspension step, then PM2 buffer is added. After the addition of the PM2 buffer the sample will begin to turn blue during mixing, when the sample is completely blue complete lysis has been achieved. Colorless areas or brown clumps indicate incomplete lysis and mixing should continue until the sample is entirely blue. During the neutralization step, PM3 buffer is added and shaking the tube vigorously will begin to turn the sample back to a neutral color. Once the sample is completely colorless, neutralization is complete. Due to the high efficiency of the Midi Column, the entire procedure can be completed in 80 minutes without ultracentrifuges, HPLC, or other toxic reagents. The purified plasmid DNA is suitable for transfection, sequencing reactions, ligation, PCR, in-vitro transcription, microinjection, restriction enzyme digestion, and gene gun.
