Which solvent is best to uniformly dissolve dioxin (HxCDD and TCDD) , causing lesser toxicity to the bacteria utilizing dioxin as a carbon source? Tween 80, Tween 20 or DMSO ?
Usually for screening for an organism capable of degrading dioxin (IN VITRO) , it is cultured in a culture medium where dioxin is dissolved in a small quantity of organic solvent, that is DMSO (1%). If the % is not higher than 1, there are no concerns about toxicity. Moreover, the organic solvent utilised can affects the degree of bacteria metabolism was strongly dependent on the solvent used, if you are interested in soil remediation maybe this review could help
regards
stefania
www.ivl.se/download/18.../B1993.pdf
Dear Dony,
Please try the following well established procedure:
Bacterial strains were routinely cultivated in a carbon-free mineral medium (CFMM) supplemented with dioxin at a concentration of 0.05% (wt/vol) at 30°C. Dioxin was added to CFMM from stock solutions (0.1 g/ml) dissolved in dimethyl sulfoxide and sterilized by filtration. Ten milliliters of precultures grown on CFMM supplemented with 0.05% (wt/vol) dioxin, was inoculated into 1 liter of the same fresh medium in 5-liter flasks and cultivated at 30°C for another 3 and 2 days, respectively. When the optical density at 600 nm reached 1.5, cells were harvested by centrifugation, washed twice with 50 mM phosphate buffer (pH 7.4), and resuspended in a reduced volume of the same buffer (an optical density at 600 nm of approximately 20).
Best wishes,
Ilya
Very valuable answers by Dr. Squadrone and Dr. Tayrlov. I would avoid Tween 80 or Tween 20, because of their bactericidal nature.
Dr. Squadrone, Dr. Tayrlov, Dr Starnes and Dr Loganathan thank you all for your response . Really appreciate it.
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