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Main sol. 1005

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Internal Solution-ID: S2051

Equipment needed: Autoclave Filter Serum bottle Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.78 g 0.772 4.43
KH2PO4 0.75 g 0.742 5.451
Na2-EDTA 0.04 g 0.04 0.118
FeSO4 x 7 H2O 0.01 g 0.01 0.036
MgSO4 x 7 H2O 0.25 g 0.247 1.003
CaCl2 x 2 H2O 0.03 g 0.03 0.202
Trace element solution 10.00 ml - -
NaCl 0.20 g 0.198 3.385
NH4Cl 0.50 g 0.495 9.246
Na2SO4 2.80 g 2.77 19.498
Na-acetate 0.15 g 0.148 1.809
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.247 1.408
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except vitamins and cysteine), adjust pH to 5.5 with 10 N H2SO4 and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml of medium in 50 ml bottles) and autoclave. Prior to inoculation add vitamins (sterilized by filtration) and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 5.5 - 6.0, if necessary.
2 After inoculation pressurize vials to 0.5 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
1005 THERMODESULFOBIUM MEDIUM