Plasmid isolated with traditional purification procedures normally contain high levels of endotoxins (also known as lipopolysaccharides or LPS) that can significantly interfere with transfection experiments downstream. The E.Z.N.A.® Endo-Free Plasmid Mini Kit II 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 condition, 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)
For Research Use Only. Not for use in diagnostic procedures.
|Downstream Application||Sensitive cell line transfection; gene therapy etc.|
|Starting material||5- 15 mL mL LB culture with OD600 between 2 and 3; or equivalent|
|Plasmid type||High-copy, low-copy, cosmid DNA|
|Processing mode||Manual, centrifugation|
|Throughput||1 - 24|
|DNA binding technology||Silica Mini Spin Column II (2x capacity of mini spin column)|
|Lysate clearance method||Centrifugation|
|Processing time||<30 minutes|
|Yield||40-70 µg for high copy-number; 1-15 µg for low copy-number|
|Special note||Includes endotoxin removal step|
Protocol and Resources
Product Documentation & Literature
Figure 1 Title
Figure 1. Quality and yield of plasmid DNA purified using the E.Z.N.A.® Endo-Free Plasmid Mini Kit. Plasmid DNA was isolated from 3 mL of three different bacterial cultures. DNA was eluted in 100 µL endotoxin-free elution buffer and quantitated using Thermo Fisher NanoDrop® 2000 and dynamic turbidity method.