EndoFree Plasmid Maxi Kit (10)

This anion-exchange-based kit from QIAGEN provides endotoxin-free plasmid or cosmid DNA purification suitable for advanced transfection-grade applications, including gene therapy research and transfection of sensitive cells. The purification process integrates an efficient endotoxin-removal step, eliminating the need for additional extractions or affinity columns to remove lipopolysaccharides.

Bacterial lysates are cleared by filtration using QIAfilter Maxi Cartridges, which replace centrifugation following alkaline lysis and can reduce plasmid-purification time by up to one hour. Plasmid DNA is then purified on gravity-flow QIAGEN-tips containing a unique anion-exchange resin developed exclusively for nucleic acid purification, giving DNA purity equivalent or superior to two successive rounds of CsCl gradient centrifugation, with endotoxin levels of <0.04 EU/µg DNA. Yields of up to 500 µg are achieved from culture.

The kit accommodates high-copy, low-copy, and cosmid DNA from 100–250 ml culture volumes, processed manually by gravity flow in 125 min per run. Buffer ER in the kit has been updated to exclude Triton X-114, ensuring REACH compliance and lower endotoxin levels in line with FDA vaccine standards, while reducing incubation time from 30 minutes on ice to 5 minutes at room temperature. The entire system avoids toxic substances such as phenol, chloroform, ethidium bromide, and CsCl. The endotoxin-free DNA obtained is highly suited for reproducible and reliable transfection in sensitive cell lines.