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  • Removing endotoxin from plasmid samples by Triton X-114 isothermal extraction.

Removing endotoxin from plasmid samples by Triton X-114 isothermal extraction.

Analytical biochemistry (2012-03-01)
Rui Ma, Jun Zhao, Hui-Cong Du, Shuang Tian, Li-Wen Li
ABSTRACT

Triton X-114 temperature transition extraction has been considered to be a simple and cost-effective strategy to eliminate endotoxin from plasmid preparations. However, a repeated cooling-heating process may promote the degradation of plasmid DNA. Based on the finding that the cloud point of Triton X-114 solution increases substantially in the presence of small amounts of sodium dodecyl sulfate (SDS) and that electrolytes decrease the cloud point of Triton X-114-SDS solution drastically, we designed a Triton X-114 isothermal extraction method for removing endotoxin from plasmid samples and found that it has the same endotoxin removal efficiency when compared with the temperature transition extraction method.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triton X-405 solution, 70% in H2O
Sigma-Aldrich
Triton X-114, laboratory grade
Sigma-Aldrich
Triton X-165 solution, Non-ionic, CMC-0.43 mM, suitable for electrophoresis, cloud point >100 °C
Sigma-Aldrich
IGEPAL® CA-630, viscous liquid
Sigma-Aldrich
IGEPAL® CA-630, for molecular biology
Supelco
IGEPAL® CA-630
Sigma-Aldrich
Triton X-114
Sigma-Aldrich
IGEPAL® CA-720, average Mn ~735