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Main sol. 1604

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Internal Solution-ID: S3344

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ferric citrate monohydrate 2.40 g 2.374 9.028
(NH4)2SO4 0.20 g 0.198 1.497
KH2PO4 3.00 g 2.967 21.805
MgSO4 x 7 H2O 0.50 g 0.495 2.007
CaCl2 x 2 H2O 0.25 g 0.247 1.682
Yeast extract
(OXOID) 		0.50 g 0.495 -
Allen's trace element solution 10.00 ml - -
FeCl2 x 4 H2O 0.26 g 0.257 1.294
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.04 g 0.04 0.225
Distilled water 1000.00 ml - -
1 First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except ferrous chloride, vitamins and cysteine. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add ferrous chloride (dissolved in 0.1 N HCl), vitamins (sterilized by filtration) and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to inoculation check pH and adjust to 6.0, if necessary.
1604 ATHALASSOTOGA MEDIUM