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Endotoxin Free Maxi Kit PLUS (PME10+/PME25+)
The GeneaidTM Maxi Plasmid Kit PLUS uses pre-packed anion-exchange resin columns to purify plasmid DNA from 200-1600 ml of cultured bacterial cells. This kit includes an increased volume of reagents for processing 3-6 g bacteria cell pellets. 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 in 100 minutes 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.
Quick Protocol

 

 

 

Advantages (Cat. # PME10+, PME25+)

  • Includes an increased volume of reagents for processing 3-6 g bacteria cell pellets
  • High Yield: 1.0 mg of pure transfection grade plasmid DNA from 300 ml cultured bacterial cells
  • Efficient endotoxin removal (<0.1 EU/µg DNA) verified by LAL
  • Large Sample Volume: 200-1600 ml of cultured bacterial cells
  • Format: plasmid maxiprep anion-exchange resin column
  • Elution Volume: 500 µl - 2 ml
  • Operation time: within 100 minutes
  • 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
  • PMC Buffer
  • PEL Buffer
  • RNase A
  • Plasmid Maxi Columns

Quality Control

The quality of the Geneaid™ Maxi Plasmid Kit PLUS (Endotoxin Free) is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Plasmid DNA from a 300 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the Geneaid™ Maxi Plasmid Kit. Following the purification process, a yield of more than 900 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid (1 µg) is analyzed by electrophoresis.

Plasmid Maxi Kit Endotoxin Free Funtional Test Data

Figure 1. Plasmid DNA was extracted using the Geneaid™ Maxi Plasmid Kit (Endotoxin Free). The purified supercoiled plasmid DNA [300 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml, OD600 = 4.0)], was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder
Sample
DNA Conc.
260/280
260/230
Total Yield
Test 11012.2 µg/ml1.872.251.0 mg
Test 21015.10 µg/ml1.872.291.0 mg
Plasmid DNA Midi and Maxi Kits Specifications
Specifications
Cat. #PIF025PIFE25PI025PIE25PM010/25PME10/25
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 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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