The I-Blue MINI Plasmid Kit was designed for rapid isolation of plasmid DNA from 1-7 mL of cultured bacterial cells. I-Blue 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 and RNA contaminants. Typical yields are 20-35 µg for high-copy number plasmid or 3-10 µg for low-copy number plasmid from 4 mL of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in restriction enzyme digestion, ligation, PCR, and sequencing reactions.

Advantages

• Sample: 1-7 mL of cultured bacterial cells
• Yield: Up to 50 µg of pure plasmid DNA
• Format: Plasmid spin column
• Operation Time: Within 15 minutes
• Elution Volume: 30-100 µL
• Kit Storage: Dry at room temperature (15-25°C) for up to 1 year; PD1 and RNase A mixture should be stored at 2-8°C for up to 6 months

Quality Control
The quality of the I-Blue MINI Plasmid Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4 mL overnight E. coli (DH5α) culture containing plasmid pBluescript (A600 > 2 U/mL). Following the purification process, a yield of more than 20 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.