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Geneaid™ Midi Plasmid Kit (PI025)

DNA Extraction/Plasmid DNA Purification

The Geneaid™ Midi Plasmid Kit uses pre-packed anion-exchange resin columns to purify plasmid DNA from 50-300 ml of cultured bacterial cells. TrueBlue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. Using an efficient gravity-flow procedure, plasmid DNA is bound to the column and contaminants are efficiently removed. The purified plasmid DNA is eluted then precipitated with isopropanol for desalting. The entire procedure can be completed without ultracentrifuges, HPLC or other toxic reagents and the purified plasmid DNA is suitable for transfection, sequencing reactions, ligation, PCR, in-vitro transcription, microinjection, restriction enzyme digestion and gene gun.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # PI025)

  • High Yield: 350 µg of pure transfection grade plasmid DNA from 100 ml of cultured bacterial cells
  • Sample Volume: 50-300 ml of cultured bacterial cells
  • Format: plasmid midi anion-exchange resin column
  • Elution Volume: 500 µl - 2 ml
  • Operation time: within 80 minutes (only 50 minutes when combined with the Presto™ Plasmid Concentrator)
  • Storage: dry at room temperature (15-25ºC)

APPLICATIONS

Transfection, Sequencing, Ligation, PCR, In-vitro Transcription, Microinjection, Restriction Enzyme Digestion, Gene Gun

COMPONENTS

  • PM1, PM2, PM3 Buffer
  • TrueBlue Lysis Buffer
  • PEQ Buffer
  • PMC Buffer
  • PEL Buffer
  • RNase A
  • Plasmid Midi Columns

QUALITY CONTROL

The quality of the Geneaid™ Midi Plasmid Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Plasmid DNA from a 100 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the Geneaid™ Midi Plasmid Kit. Following the purification process, a yield of more than 300 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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Figure 1 .Figure 1. Plasmid DNA from a 100 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (OD600 = 3.65) was purified using the Geneaid™ Midi Plasmid Kit. The purified supercoiled plasmid DNA was used in EcoRI digestion. Plasmid DNA yield was determined by spectrophotometer and analyzed by electrophoresis on a 1% agarose gel.

M = Geneaid 1 Kb DNA Ladder

 
Test
DNA Conc.
A260/280
A260/230
Yield
1
332.1 µg/ml
1.87
2.22
332.1 µg
2
345.5 µg/ml
1.87
2.25
345.5 µg
3
339.2 µg/ml
1.87
2.24
339.2 µg

Plasmid DNA Midi and Maxi Kits 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

RELATED PRODUCTS

 
Product NameRXNS/VOLUMECatalogue Number
Presto™ Endotoxin Free Mini Plasmid Kit100PEH100
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Large Plasmid DNA Extraction Kit100, 300PDL100/300
High-Speed Plasmid Advance Kit (50-100 ml)25PA025
Presto™ Midi Plasmid Kitt25PIF025
Presto™ Midi Plasmid Kit (Endotoxin Free)25PIFE25
Geneaid™ Midi Plasmid Kit25PI025
Geneaid™ Midi Plasmid Kit (Endotoxin Free)25PIE25
Presto™ Plasmid DNA Concentration Kit25, 50, 100PC0250/500/1000
Geneaid™ Maxi Plasmid Kit10, 25PM010/25
Geneaid™ Maxi Plasmid Kit (Endotoxin Free)10, 25PME10/25
Presto™ 96 Well Plasmid Kit4 x 96, 10 x 9696PDC04/10
96PDV04/10
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