Midiprep Plasmid DNA Kit (PIF025)

Midi Plasmid Kit
Midiprep Plasmid DNA Kit (PIF025)

The Presto™ Midi Plasmid Kit was designed for Plasmid DNA Purification 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 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 80 minutes without ultracentrifuges, HPLC or other toxic reagents.  

Advantages (Cat. # PIF025)
  • High Yield: >400 µg of pure transfection grade plasmid DNA from 100 ml of cultured bacterial cells
  • 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 (membrane optimized for plasmid DNA purification)
  • Elution Volume: 500 µl - 2 ml
  • Storage: dry at room temperature (15-25°C) for up to 2 years without showing any reduction in performance
  • Operation time: within 80 minutes (only 40 minutes when combined with the Presto™ Plasmid Concentrator)
Applications
 
Transfection, Sequencing, Ligation, PCR, In-vitro Transcription, Microinjection, Restriction Enzyme Digestion, Gene Gun
 
Components
  • PM1, PM2, PM3 Buffers
  • TrueBlue Lysis Buffer
  • PEQ Buffer
  • PW 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 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.
 
Presto™ Midi Plasmid Kit Functional Test Data
Brand
Test
260/280
260/230
Yield
Average
Geneaid
(Lane 1, 2)
50 ml
1.85
2.25
273.00 µg
265.90 µg 
50 ml
1.87
2.23
258.80 µg
100 ml
1.87
2.30
373.20 µg
391.20 µg
100 ml
1.87
2.14
409.20 µg
Competitor
(Lane 3, 4)
50 ml
1.85
2.26
121.10 µg
131.30 µg
50 ml
1.85
2.29
141.50 µg
100 ml
1.86
2.32
285.80 µg
287.85 µg
100 ml
1.87
2.33
289.90 µg
Figure 1. Plasmid DNA was extracted using the Presto™ Midi Plasmid Kit (lane 1,2) and the equivalent competitor's plasmid midi kit (lane 3,4). The purified super coiled 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 (50 ml), Lane 2: Presto™ Midi Plasmid Kit (100 ml), Lane 3: Equivalent Competitor Kit (50 ml), Lane 4: Equivalent Competitor Kit (100 ml)
Column Types Included With the Presto™ Midi Plasmid Kit
Filter Column               
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 exhange resin column is used for plasmid DNA  binding, washing and eluting.
TrueBlue Lysis Buffer Optional Color Indicator
Cell Lysis
Neutralization
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.
Insufficient Mixing
Correct Mixing
Insufficient Mixing
Correct Mixing
Plasmid DNA Midi and Maxi Kits Specifications 
Specifications
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 500 µg 500 µg 800 µg 800 µg 1.5 mg 1.5 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 200-500 µg 200-500 µg 200-800 µg 200-800 µg 1-1.5 mg 1-1.5 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 <120 minutes <120 minutes <180 minutes <180 minutes
Related Plasmid DNA Purification Products
Product Name RXNS Catalogue Number
Presto™ Endotoxin Free Mini Plasmid Kit 100 PEH100
100, 300
PDH100/300
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PD100/300
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PDL100/300
25
PA025
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PIF025
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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