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.
- 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)
- 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)
Figure 1. Plasmid DNA was extracted using both the Presto™ Midi Plasmid Kit Endotoxin Free (lane 1,2) and the equivalent competitor's plasmid midi kit (lane 3,4). The purified supercoiled plasmid DNA [50 ml and 100 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml, OD600 = 3.8)], was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel.
M = Geneaid 1 Kb DNA Ladder, Lane 1: Presto™ Midi Plasmid Kit Endotoxin Free (50 ml), Lane 2: Presto™ Midi Plasmid Kit Endotoxin Free (100 ml), Lane 3: Equivalent Competitor Kit (50 ml), Lane 4: Equivalent Competitor Kit (100 ml)
Figure 2. This centrifuge midi plasmid spin column is used to filter the neutralized colorless mixture and remove contaminants to facilitate DNA binding to the white membrane of the Plasmid Midi Column.
|Plasmid Midi Column|
|Figure 3. This gravity flow plasmid midi anion exchange resin column is used for binding, washing and eluting plasmid DNA.|
Figure 4. TrueBlue Lysis Buffer is an optional color indicator included with this Plasmid Kit to eliminate common handling errors and ensure efficient cell lysis and neutralization each and every time.
|Binding Capacity||350 µg||350 µg||350 µg||350 µg||1.0 mg||1.0 mg|
|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|
|Product Name||RXNS||Catalogue Number|
|Presto™ Endotoxin Free Mini Plasmid Kit||100||PEH100|
|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|