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Main sol. 284

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Internal Solution-ID: S552

Equipment needed: Filter Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.14 g 0.138 0.794
CaCl2 x 2 H2O 0.14 g 0.138 0.941
NH4Cl 0.25 g 0.247 4.618
MgSO4 x 7 H2O 3.40 g 3.36 13.631
MgCl2 x 6 H2O 4.10 g 4.051 19.928
KCl 0.33 g 0.326 4.374
NiCl2 x 6 H2O
(0.1% w/v) 		0.50 ml 4.9e-4 0.004
NaCl 30.00 g 29.644 507.26
Fe(NH4)2(SO4)2 x 6 H2O 0.01 g 0.01 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.50 g 1.482 13.985
Yeast extract 1.00 g 0.988 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.494 2.813
Na2S x 9 H2O 0.50 g 0.494 2.057
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, yeast extract, vitamins, cysteine and sulfide. 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 to 30% of their volume and autoclave. Add vitamins (sterilized by filtration), yeast extract, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use check pH of completed medium and adjust to 7.0 - 7.2, if necessary.
2 After inoculation add sterile 80% N2 and 20% CO2 gas mixture to 1 bar overpressure.
284 KOSMOTOGA MEDIUM