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Main sol. 282b

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Internal Solution-ID: S550

Equipment needed: Autoclave Filter Gassing station Hungate tubes
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
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.00 g 0.988 11.763
Na2S2O3 x 5 H2O 1.00 g 0.988 3.981
Wolin's vitamin solution (10x) 1.00 ml - -
DL-Dithiothreitol
(DTT) 		1.00 g 0.988 6.404
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, sodium thiosulfate, vitamins, and DTT. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, then dispense medium into anoxic tubes to 30% of their volume under 80% H2 and 20% CO2 gas atmosphere and autoclave. Add thiosulfate, vitamins and DTT from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration. Prior to use check pH of completed medium and adjust to 6.0, if necessary.
2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.
282b DESULFUROBACTERIUM HR11 MEDIUM