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Midiprep Plasmid DNA Kit (PIF025)

DNA Extraction/Plasmid DNA Purification

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 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.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # PIF025)

  • High Yield: 350 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)
  • Operation time: within 60 minutes (only 30 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
  • 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 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™ Mini Plasmid Kit Functional Test Data

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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
265.90 µg
100 ml
1.87
2.30
373.20 µg
391.20 µg
100 ml
1.87
2.14
409.20 µg
391.20 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
131.30 µg
100 ml
1.86
2.32
285.80 µg
287.85 µg
100 ml
1.87
2.33
289.90 µg
287.85 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

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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.

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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
  • *
    Insufficient Mixing
  • *
    Correct Mixing
Neutralization
  • *
    Insufficient Mixing
  • *
    Correct Mixing
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.

Plasmid DNA Kit Specifications

 
SpecificationsPresto™ Midi Plasmid KitPresto™ Midi Plasmid Kit Endotoxin FreeGeneaid™ Midi Plasmid KitGeneaid™ Midi Plasmid Kit Endotoxin FreeGeneaid™ Maxi Plasmid KitGeneaid™ Maxi Plasmid Kit Endotoxin Free
Cat. #PIF025PIFE25PI025PIE25PM010/25PME10/25
FormatAnion-Exchange Gravity FlowAnion-Exchange Gravity FlowAnion-Exchange Gravity FlowAnion-Exchange Gravity FlowAnion-Exchange Gravity FlowAnion-Exchange Gravity Flow
Binding Capacity350 µg350 µg350 µg350 µg1.0 mg1.0 mg
Filter ColumnYesYesN/AN/AN/AN/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 Input50-150 ml50-150 ml50-300 ml50-300 ml200-1600 ml200-1600 ml
Plasmid Size1-20 kb1-20 kb1-20 kb1-20 kb1-20 kb1-20 kb
Typical Yield350 µg350 µg350 µg350 µg1.0 mg1.0 mg
Elution Volume500 µl-2 ml500 µl-2 ml500 µl-2 ml500 µl-2 ml500 µl-2 ml500 µl-2 ml
Operation Time<80 minutes<80 minutes<80 minutes<80 minutes<100 minutes<100 minutes

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