The I-Blue Mini Plasmid Endo-Free Kit was designed for transfection grade plasmid DNA purification from 1-5ml of cultured E.coli cells using silica-membrane spin columns. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. In addition, an efficient endotoxin removal step is integrated in the procedure and endotoxins are precipitated in a proprietary reagent. 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. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 50 minutes. The purified endotoxin free plasmid DNA is ready-for-use in transfection, restriction enzyme digestion, ligation, PCR, and sequencing reactions.
The quality of the I-Blue Mini Plasmid Endo-Free Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600>2U/mL). Following the purification process, a yield of more than 25µg is obtained and the ratio of A260/A280 is between 1.8-2.0. The purified plasmid DNA (1µg) is used in EcoRI digestion and analyzed by electrophoresis.