E.Z.N.A.® Endo-free Plasmid DNA Mini Kit I

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Omega Biotek E.Z.N.A.® Endo-free Plasmid DNA Mini Kit I

Isolate low endotoxin (<1 EU/µg) plasmid DNA from up to 5 mL bacterial cultures using spin mini columns

Plasmid isolated with traditional purification procedures normally contains high levels of endotoxins (also known as lipopolysaccharides or LPS) that can significantly interfere with transfection experiments downstream. Up to 5 mL of overnight cultures can be used for high copy number plasmids and 10 mL for low copy number.

The E.Z.N.A.® Endo-Free Plasmid DNA Mini Kit I integrates an efficient endotoxin removal step into the plasmid purification procedure to produce high-quality transfection grade (<1 EU/µg) plasmid for efficient transfection. The bacterial cells are lysed using the alkaline-SDS lysis method. The cleared cell lysate is then treated with ETR reagent to efficiently remove the endotoxins. After adjusting the binding conditions, the cell lysate is applied into the HiBind® DNA column and purified DNA is eluted from the column membrane.

  • Rapid – 30 minutes or less
  • Safe – No phenol/chloroform extractions
  • Versatile – Spin and vacuum formats available
  • High-quality – Endotoxins efficiently reduced (<1 EU/µg)
Downstream ApplicationSensitive cell line transfection; gene therapy etc.
Starting Material1-5 mL LB culture with OD600 between 2 and 3; or equivalent
Plasmid TypeHigh-copy, low-copy, cosmid DNA
Processing modeManual, centrifugation
Throughput1 – 24
DNA binding technologySilica mini spin column
Lysate clearing methodCentrifugation
Processing Time<30 minutes
Yield15-25 µg for high copy-number; 0.1-5 µg for low copy-number
OtherIncludes endotoxin removal step