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Main sol. 784a

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Internal Solution-ID: S1590

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.2 g 0.2 1.467
MgCl2 x 6 H2O 0.1 g 0.1 0.491
NH4Cl 1.0 g 0.998 18.658
KCl 0.2 g 0.2 2.677
NaCl 60.0 g 59.88 1024.64
Trace element solution SL-11 1.0 ml - -
Na-acetate 0.2 g 0.2 2.433
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
Na2CO3 68.0 g 67.864 640.301
NaHCO3 38.0 g 37.924 451.44
Wolin's vitamin solution (10x) 1.0 ml - -
Na2S x 9 H2O 1.0 g 0.998 4.155
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, bicarbonate, vitamins and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve carbonates and sulfide while gassing the head space only. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add vitamins from an anoxic stock solution sterilized by filtration. Adjust pH of the complete medium to 9.0 - 9.5.
784a METHANOCALCULUS NATRONOPHILUS MEDIUM