I am trying to nucleofect/electroporate my T cell clones. These clones seem to be very sensitive to endotoxin in the plasmid preparation. I wonder if someone could give me some recommendation of the best endotoxin-free kit they have used. Thank you guys!
I had good experience with two endotoxin free plasmid preps. Endo toxin free Qiagen maxi-prep is best so far for plasmid preparation. I can't say that it will have zero endotoxin levels. However, the levels are far below which has significantly little interference with transfection procedures. In my experimental studies, I tried and confirmed it. A comparatively inexpensive one is from Sigma (GenElute™ Endotoxin-free Plasmid Midiprep Kit). But it has limitations are, the plasmid concentrations are far below. Sometimes, a diluted plasmid (in water) may not allow efficient transfections. I haven't tested GeneJET Endo-Free Plasmid Maxiprep Kit from Thermo.
I have always used the Qiagen maxi or mini prep kits and it always worked better than with different other kits and generated every times excellent results with high purity and satisfactory concentrations.
PureLink endotoxin-free kits. Ultraclean plasmid, up to Giga scale: https://www.thermofisher.com/order/catalog/product/A31232
In case you didn't know, there is an automated kit on the market. We now offer an automated instrument that does endotoxin free Maxiprep and consistently gives you 1mg of plasmid DNA from a 5g cell culture wet weight. Resin is anion exchange based.
Info: https://phynexus.com/phyprep/
App NoteExperiment Findings The Biotage® PhyPrep system and fully automated maxiprep pla...
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