HIGHLY PURE PLASMID DNA PREPARATIONS
The present disclosure generally relates to highly pure plasmid compositions having low, or undetectable, levels of colanic acid and other contaminants. The compositions described herein have a range of uses, including diverse applications in the field of bioterrorism, environmental science, food science, forensics, molecular biology, and health and medicine.
1 . A gram negative bacterial plasmid composition comprising gram negative bacterial plasmid DNA and less than about 0.1 mg of colanic acid per mg of gram negative bacterial plasmid DNA.
2 . The composition of claim 1 , wherein the composition comprises less than about 0.05 mg of colanic acid per mg of gram negative bacterial plasmid DNA.
3 . The composition of claim 1 , wherein the composition comprises no detectable colanic acid.
4 . The composition of claim 1 , wherein the composition comprises no detectable colanic acid, as measured by a bicinchoninic acid (BCA) assay.
5 . The composition of claim 1 , wherein the composition further comprises less than about 0.1 mg uronic acid per mg of gram negative bacterial plasmid DNA.
6 . The composition of claim 1 , wherein the composition further comprises less than about 0.05 mg uronic acid per mg of gram negative bacterial plasmid DNA.
7 . The composition of claim 1 , wherein the composition comprises no detectable uronic acid.
8 . The composition of claim 1 , wherein the composition comprises no detectable uronic acid, as measured by a uronic acid assay.
9 . The composition of claim 1 , wherein the composition further comprises less than about 0.1 mg fucose per mg of gram negative bacterial plasmid DNA.
10 . The composition of claim 1 , wherein the composition further comprises less than about 0.05 mg fucose per mg of gram negative bacterial plasmid DNA.
11 . The composition of claim 1 , wherein the composition comprises no detectable fucose.
12 . The composition of claim 1 , wherein the composition comprises no detectable fucose, as measured by a fucose assay.
13 . The composition of claim 1 , wherein the composition comprises no visually detectable polysaccharides when combined with a polysaccharide-selective labeling agent.
14 . The composition of claim 13 wherein the polysaccharide-selective labeling agent is (4,6-dichlorotriazinyl)aminofluorescein (DTAF).
15 . A gram negative bacterial plasmid composition comprising gram negative bacterial plasmid DNA, less than about 0.1 mg of colanic acid per mg of gram negative bacterial plasmid DNA, and less than about 0.1 mg uronic acid per mg of gram negative bacterial plasmid DNA.
16 . The composition of claim 15 wherein the composition further comprises less than about 0.1 mg fucose per mg of gram negative bacterial plasmid DNA.
17 . The composition of claim 16 wherein the composition comprises no detectable colanic acid, uronic acid, and fucose.
18 . A gram negative bacterial plasmid composition comprising gram negative bacterial plasmid DNA, less than about 0.05 mg of colanic acid per mg of gram negative bacterial plasmid DNA, and less than about 0.05 mg fucose acid per mg of gram negative bacterial plasmid DNA.
19 . The composition of claim 15 wherein the composition further comprises less than about 0.05 mg uronic acid per mg of gram negative bacterial plasmid DNA.
20 . The composition of claim 19 wherein the composition further comprises no detectable colanic acid, fucose, uronic acid chromosomal or genomic DNA, RNA, protein, and/or endotoxin contaminants.