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Main sol. 1570

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Internal Solution-ID: S3255

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.528 505.263
CaCl2 x 2 H2O 0.15 g 0.148 1.004
MgCl2 x 6 H2O 3.05 g 3.002 14.766
KCl 0.33 g 0.325 4.357
(NH4)2SO4 0.66 g 0.65 4.916
Modified Wolin's mineral solution 10.00 ml - -
Phosphate buffer 5.00 ml - -
Na2S2O3 x 5 H2O 1.50 g 1.476 5.949
Na2CO3 1.00 g 0.984 9.286
Wolin's vitamin solution (10x) 1.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except phosphate buffer, thiosulfate, carbonate and vitamins) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials up to a volume of 20% and autoclave. Add phosphate buffer, thiosulfate, carbonate, and vitamin solution to the autoclaved medium from sterile anoxic stock solutions prepared under 100% N2 gas. Phosphate buffer, stock solutions of vitamins and thiosulfate are sterilized by filtration and stored under N2, whereas the solution of carbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust pH of the complete medium to 6.7, if necessary.
2 After inoculation pressurize vessels to 0.5 bar overpressure with sterile 80% N2 and 20% CO2 gas mixture and add sterile air in an amount that is equivalent to a volume of 20% of the headspace.
1570 THIOGRANUM MEDIUM