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Main sol. 399

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Internal Solution-ID: S812

Equipment needed: Autoclave Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.336 4.507
MgCl2 x 6 H2O 4.00 g 3.953 19.442
MgSO4 x 7 H2O 3.45 g 3.409 13.831
NH4Cl 0.25 g 0.247 4.618
CaCl2 x 2 H2O 0.14 g 0.138 0.941
K2HPO4 0.14 g 0.138 0.794
NaCl 18.00 g 17.787 304.356
Yeast extract
(OXOID) 		0.50 g 0.494 -
Na-L-lactate 1.50 g 1.482 -
Fe(NH4)2(SO4)2 x 7 H2O
(0.1% w/v) 		2.00 ml 0.002 0.005
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 3.00 g 2.964 35.288
Na2S x 9 H2O 0.50 g 0.494 2.057
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and adjust pH to 6.9, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into serum vials (e.g., 20 ml per 100 ml bottle), seal and autoclave. Before use reduce the medium with sodium sulfide from a sterile anoxic stock solution prepared under 100% N2 gas.
2 After inoculation, add 2 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture.
399 ARCHAEOGLOBUS MEDIUM