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Main sol. 206

Internal Solution-ID: S439

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.0 g 3 21.121
NH4Cl 1.0 g 1 18.695
MgCl2 x 6 H2O 0.2 g 0.2 0.984
KH2PO4 0.3 g 0.3 2.205
Na2HPO4 x 12 H2O 2.0 g 2 14.089
Trace element solution 10.0 ml - -
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
1.5 ml 0.002 0.005
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 -
Yeast extract 1.0 g 1 -
Na-L-lactate 4.0 g 4 -
Wolin's vitamin solution 5.0 ml - -
Na2S x 9 H2O 0.5 g 0.5 2.082
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except yeast extract, lactate, vitamins and sulfide) and sparge medium for 30 - 45 min with 100% N2 gas to make it anoxic. Distribute under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add yeast extract, lactate, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Before use, neutralize the sodium sulfide solution by dropwise addition of 2 N H2SO4 (see medium 29). Adjust pH of complete medium to 6.8 - 7.0.
206 THERMODESULFOBACTERIUM MEDIUM