Presto™ Midi Plasmid Kit (Endotoxin Free)
DNA Extraction/Plasmid DNA Purification
The Presto™ Midi Plasmid Kit Endotoxin Free was designed for Plasmid DNA Purification and endotoxin removal from 50-150 ml of cultured bacterial cells using an efficient plasmid midiprep system. For processing smaller volumes, the Presto™ Mini Plasmid Kit is also available. Both plasmid kits include TrueBlue Lysis Buffer (an optional color indicator) to prevent common handling errors, while ensuring efficient cell lysis and neutralization. The Presto™ Midi Plasmid Kit Endotoxin Free 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 PMC 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 80 minutes without ultracentrifuges, HPLC or other toxic reagents.
Specifications (Cat. # PIFE25)
- High Yield: 350 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
- Convenient: TrueBlue Lysis Buffer is an optional color indicator included with this plasmid kit in order to reduce common handling errors, while ensuring efficient cell lysis and neutralization and maximum plasmid product yield so you can feel confident with each and every plasmid prep.
- Sample Volume: 50-150 ml of cultured bacterial cells
- Format: Plasmid Midi Anion-Exchange Resin Columns and Filter Columns (membranes optimized for plasmid DNA purification)
- Elution Volume: 500 µl – 2 ml
- Storage: dry at room temperature (15-25ºC)
- Operation time: within 60 minutes (only 30 minutes when combined with the Presto™ Plasmid Concentrator)
- Transfection,
- Sequencing,
- Ligation,
- PCR,
- In-vitro Transcription,
- Microinjection,
- Restriction Enzyme Digestion,
- Gene Gun
Plasmid DNA Kit Specifications
| Specifications | Presto™ Midi Plasmid Kit | Presto™ Midi Plasmid Kit Endotoxin Free | Geneaid™ Midi Plasmid Kit | Geneaid™ Midi Plasmid Kit Endotoxin Free | Geneaid™ Maxi Plasmid Kit | Geneaid™ Maxi Plasmid Kit Endotoxin Free | ||
| Cat. # | PIF025 | PIFE25 | PI025 | PIE25 | PM010/25 | PME10/25 | ||
| Format | Anion-Exchange Gravity Flow | Anion-Exchange Gravity Flow | Anion-Exchange Gravity Flow | Anion-Exchange Gravity Flow | Anion-Exchange Gravity Flow | Anion-Exchange Gravity Flow | ||
| Binding Capacity | 350 µg | 350 µg | 350 µg | 350 µg | 1.0 mg | 1.0 mg | ||
| Filter Column | Yes | Yes | N/A | N/A | N/A | N/A | ||
| Endotoxin Level | <0.15 EU/µg | <0.10 EU/µg | <0.15 EU/µg | <0.10 EU/µg | <0.15 EU/µg | <0.10 EU/µg | ||
| Culture Input | 50-150 ml | 50-150 ml | 50-300 ml | 50-300 ml | 200-1600 ml | 200-1600 ml | ||
| Plasmid Size | 1-20 kb | 1-20 kb | 1-20 kb | 1-20 kb | 1-20 kb | 1-20 kb | ||
| Typical Yield | 350 µg | 350 µg | 350 µg | 350 µg | 1.0 mg | 1.0 mg | ||
| Elution Volume | 500 µl-2 ml | 500 µl-2 ml | 500 µl-2 ml | 500 µl-2 ml | 500 µl-2 ml | 500 µl-2 ml | ||
| Operation Time | <80 minutes | <80 minutes | <80 minutes | <80 minutes | <100 minutes | <100 minutes |
Components
- PM1, PM2, PM3 Buffers
- PER Buffer (Endotoxin Removal Buffer)
- TrueBlue Lysis Buffer
- PEQ Buffer
- PMC Buffer
- PEL Buffer
- RNase A
- Filter Columns (yellow spin columns)
- Plasmid Midi Columns (anion exchange white resin columns)
Quality Control
The quality of the Presto™ 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 Presto™ Midi Plasmid Kit. Following the purification process, a yield of more than 200 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.
Column Types Included With the Presto™ Midi Plasmid Kit
Filter Column
Figure 2.This centrifuge filter spin column is used to remove the contaminants such as cell debris and genomic DNA from the neutralized colorless mixture to facilitate plasmid DNA binding to the Plasmid Midi Column.
Plasmid Midi Column
Figure 3.This gravity flow plasmid midi anion exhange resin column is used for plasmid DNA binding, washing and eluting.
| RELATED PRODUCTS | ||
|---|---|---|
| Product Name | RXNS/VOLUME | Catalogue Number |
| Presto™ Endotoxin Free Mini Plasmid Kit | 100 | PEH100 |
| Presto™ Mini Plasmid Kit | 100, 300 | PDH100/300 |
| High-Speed Plasmid Mini Kit | 100, 300 | PD100/300 |
| Large Plasmid DNA Extraction Kit | 100, 300 | PDL100/300 |
| Midiprep Spin Column Plasmid Kit | 25 | PA025 |
| Presto™ Midi Plasmid Kit | 25 | PIF025 |
| Presto™ Midi Plasmid Kit (Endotoxin Free) | 25 | PIFE25 |
| Geneaid™ Midi Plasmid Kit | 25 | PI025 |
| Geneaid™ Midi Plasmid Kit (Endotoxin Free) | 25 | PIE25 |
| Presto™ Plasmid DNA Concentration Kit | 25, 50, 100 | PC0250/500/1000 |
| Geneaid™ Maxi Plasmid Kit | 10, 25 | PM010/25 |
| Geneaid™ Maxi Plasmid Kit (Endotoxin Free) | 10, 25 | PME10/25 |
| Presto™ 96 Well Plasmid Kit | 4 x 96, 10 x 96 | 96PDC04/10 96PDV04/10 |
