Endotoxin Free Maxi Kit PLUS (PME10+/PME25+)

Maxi Plasmid Kit Endotoxin Free
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 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. # PME10+, PME25+)

  • Includes an increased volume of reagents for processing 3-6 g bacteria cell pellets
  • High Yield: 1.5 mg of pure transfection grade plasmid DNA from 400 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 2.5 hours
  • Storage: dry at room temperature (15-25ºC)

Applications

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

Compnents

  • PM1, PM2, PM3 Buffer
  • TrueBlue Lysis Buffer
  • PER Buffer
  • PEQ Buffer
  • PW 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 400 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 used in EcoRI digestion, and 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 [400 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml, OD600 = 4.0)], was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder
Sample
DNA Conc.
260/280
260/230
Total Yield
Test 1 1550.10 µg/ml 1.87 2.25 1.55 mg
Test 2 1540.10 µg/ml 1.87 2.29 1.54 mg
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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