Geneaid™ Midi Plasmid Kit Endotoxin Free (PIE25)

Midi Plasmid Kit Endotoxin Free
Geneaid™ Midi Plasmid Kit Endotoxin Free (PIE25)
The Geneaid™ Midi Plasmid Kit Endotoxin Free 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.

 

 

Advantages (Cat. # PIE25)

  • High Yield: >400 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
  • 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 2 hours (only 80 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
  • PER Buffer
  • PEQ Buffer
  • PW Buffer
  • PEL Buffer
  • RNase A
  • Plasmid Midi Columns

Quality Control

The quality of the Geneaid™ 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 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 400 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid DNA (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.

Plasmid Midi Kit Endotoxin Free Functional Test Data

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 Midi. 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
1 211.2 µg/ml 1.87 2.27 422.4 µg
2 216.6 µg/ml 1.87 2.27 433.2 µg
3 224.8 µg/ml 1.87 2.28 449.6 µg
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
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