The E.Z.N.A.® Endo-Free Plasmid Midi/Maxi Kits provide an efficient method for the large-scale isolation of endotoxin-free plasmid DNA. Following lysis, endotoxins are removed from the cleared lysate with the addition of ETR Solution. Plasmid DNA is then further purified by adsorbing to the HiBind silica membrane. An easy wash step is followed by the elution of the bound plasmid DNA in the elution buffer. Up to 200 µg of high-quality plasmid DNA can be purified from 15-50 mL of overnight culture (D6915), and up to 1 mg from 50-200 mL of overnight culture (D6926). Purified DNA samples are ready to be used in downstream applications such as automated fluorescent DNA sequencing, restriction enzyme digestion, transfection of mammalian cells, cloning, and PCR amplification.
- Rapid – Purification of plasmid DNA in < 45min
- Safe – No phenol/chloroform extractions
- 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||15-50 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 Midi Spin Column|
|Lysate clearance method||Centrifugation|
|Processing time||<30 minutes|
|Yield||100-250 µg for high copy-number; 10-50 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 Midi & Maxi Kits. Plasmid (PEGF-N1) DNA was isolated from 50 mL (Midi Kit)/200 mL (Maxi Kit) of LB cultures. DNA was eluted in 500 µL (Midi Kit)/2 mL (Maxi Kit) endotoxin-free elution buffer and quantitated using Thermo Fisher NanoDrop® 2000 and dynamic turbidity method.