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Main sol. 901a

Internal Solution-ID: S1854

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.324 6.066
KCl 0.33 g 0.324 4.353
KH2PO4 0.33 g 0.324 2.384
MgCl2 x 6 H2O 0.33 g 0.324 1.596
CaCl2 x 2 H2O 0.33 g 0.324 2.207
Na2SO4 2.80 g 2.753 19.383
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Yeast extract 3.00 g 2.95 -
Wolin's vitamin solution (10x) 1.00 ml - -
Neutralized sulfide solution 3% (w/v) 15.00 ml - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except yeast extract, vitamins and sulfide. Adjust pH to 4.5 with H2SO4 and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The sulfide stock solution (3% w/v) should be neutralized before use (see medium 28). The pH of the complete medium should be at 4.5.
901a THERMODESULFOBIUM ACIDIPHILUM MEDIUM