The Midi Plasmid Kit Endotoxin Free was designed for plasmid DNA purification and endotoxin removal using pre-packed anion-exchange resin midiprep columns to purify plasmid DNA from 50-300 ml of cultured bacterial cells. This endotoxin free plasmid kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. Using an efficient gravity-flow procedure, plasmid DNA in the crude lysate is bound by the plasmid midi column and the contaminants are removed with PW Buffer. The purified plasmid DNA is eluted by a high salt buffer and then precipitated with Isopropanol for desalting. The entire procedure can be completed in 2 hours without ultracentrifuges, HPLC or other toxic reagents and the purified plasmid DNA is suitable for Transfection, Sequencing Reactions, Ligation, PCR, and In-vitro Transcription, Microinjection, Restriction Enzyme Digestion and Gene Gun.
Advantages:
High Yield: >700 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: anion-exchange resin column
Operation time: within 2 hours
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
PER Buffer
PEQ Buffer
PW Buffer
PEL Buffer
RNase A
Plasmid Midi Columns
Quality Control:
The quality of the Plasmid Midi 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 (DH5?) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the Plasmid Midi Kit. Following the purification process, a yield of more than 300 ?g 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.
Codice Prodotto: PIFE25
