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EndoFree Plasmid Kits

transfection experiments by competing with DNA for free transfection reagent. Endotoxins also induce nonspecific activation of immune responses in immune cells such as macrophages and B cells, which can lead to misinterpretation of transfection results. These responses include induced synthesis of proteins and lipids such as IL-1 and prostaglandin. Overall, endotoxins represent a non-controllable variable in transfection experiment setup, influencing the outcome and reproducibility of results and making them difficult to compare and interpret. In gene therapy research, endotoxins can interfere by causing endotoxic-shock syndrome and activation of the complement cascade.
EndoFree principle

The level of endotoxin contamination in purified plasmid DNA depends on the purification method used (see table "Endotoxin levels in plasmid preparations"). Silica-slurrypurified DNA exhibits extremely high endotoxin levels. QIAGEN, QIAfilter, and HiSpeed Plasmid Kits and 2x CsCl ultracentrifugation yield very pure DNA with relatively low levels of endotoxin. EndoFree Plasmid Kits include an integrated endotoxin-removal step to yield plasmid DNA containing <0.1 EU/g plasmid DNA.

Procedure

Following clearing of the bacterial lysate using the QIAfilter Cartridge (see "QIAfilter principle" ), the Endotoxin Removal Buffer is added to the filtered lysate which is incubated on ice. The lysate is then applied directly to a QIAGEN-tip for plasmid DNA purification.
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