[Purification of plasmid DNA using anion-exchange chromatography and removal of endotoxin].
The plasmid DNA was purified using the Fractogel EMD TMAE (M) anion exchange media. The dynamic loading capacity of this media for plasmid DNA was 0.62 mg/mL. It was found that when the lysate of bacteria was incubated with either Triton X-114 or Triton X-100 before purification, the average content of endotoxin left in the purified plasmid DNA was 6.42 EU/mg or 9.50 EU/mg, respectively. These results were much lower than that without pre-incubation (67.82 EU/mg). The purification of plasmid DNA and the removal of endotoxin were achieved in one step using anion-exchange chromatography. This method is simple, fast and of low cost.
Pi
Wenhui
W
College of Animal Science and Technology, Shihezi University, Shihezi 832003, China. wzjpwh@163.com
Sun
Congjian
C
Song
Zhiqiang
Z
Ma
Lianying
L
Liu
Shuping
S
Bai
Dingping
D
Cai
Yidong
Y
Liu
Shouren
S
D000837:Anion Exchange Resins / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004731:Endotoxins / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D017830:Octoxynol / Q000737:chemistry; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification*
D000837:Anion Exchange Resins; D004731:Endotoxins; D017830:Octoxynol; D004247:DNA