Removing endotoxin from plasmids
malexe at biost1.thi.tmc.edu
Thu Aug 1 09:35:11 EST 1996
In article <msklar-3107962134520001 at msklar.cts.com>, msklar at cts.com (Mike
> Hi. I'm hoping someone can help me out. I've been trying to purify
> endotoxin free plasmid DNA for in vivo work using a variety of methods.
> I've tried using various kits and Phenol/chloroform to clean it up and
> nothing seems to work. My concentrations are still 2-5 EU/ug. Can
> anyone offer a useful protocol or method for cleaning up the DNA? I'd
> really appreciate it.
> Mike Sklar
Qiagen sells special columns that allow to isolate DNA that is essentially
endotoxin-free. Double banding in CsCl also seems to work well, although
results are not very reproducible. You may also look at:
Removal of endotoxins by affinity sorbents. Anspach FB. Hilbeck O.
Journal of Chromatography A. 711(1):81-92, 1995 Sep 8.
Bacterial lipopolysaccharide copurifies with plasmid DNA: implications
for animal models and human gene therapy. Wicks IP. Howell ML.
Human Gene Therapy. 6(3):317-23, 1995 Mar.
Lipopolysaccharide is a frequent contaminant of plasmid DNA
preparations and can be toxic to primary human cells in the presence of
Gene Therapy. 1(4):239-46, 1994 Jul.
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