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Main sol. 845

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Internal Solution-ID: S1732

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Peptone
(BD Bacto) 		5 g 4.985 -
Disodium succinate 1 g 0.997 6.152
(NH4)2SO4 1 g 0.997 7.545
MgSO4 x 7 H2O 1 g 0.997 4.045
FeCl3 x 6 H2O 2 mg 0.002 0.007
MnSO4 x H2O 2 mg 0.002 0.012
Agar
(BD Bacto) 		1 g 0.997 -
Trace element solution SL-11 1 ml - -
Seven vitamins solution 1 ml - -
Catalase
(2000 U/ml) 		1 ml - -
Distilled water 1000 ml - -
1 Dissolve ingredients except trace elements, vitamins and catalase, adjust pH to 7.2 - 7.4, bring medium to the boil and cool to room temperature under 100% N2 gas atmosphere. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. To complete the medium add trace elements, vitamins and catalase from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins and catalase should be sterilized by filtration. After inoculation, inject sterile air into the culture vessel up to about 20% of the total gas volume (i.e., 4 parts N2 and 1 part air).
845 SPIRILLUM WINOGRADSKYI MEDIUM