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Main sol. 561

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Internal Solution-ID: S1231

Equipment needed: Autoclave Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.30 g 0.296 3.965
MgCl2 x 6 H2O 2.75 g 2.709 13.327
MgSO4 x 7 H2O 3.45 g 3.399 13.79
NH4Cl 1.25 g 1.232 23.024
CaCl2 x 2 H2O 0.14 g 0.138 0.938
K2HPO4 0.14 g 0.138 0.792
NaCl 0.50 g 0.493 8.429
Modified Wolin's mineral solution 10.00 ml - -
NiCl2 x 6 H2O
(0.1% w/v) 		5.00 ml 0.005 0.038
Na2WO4 x 2 H2O
(0.1% w/v) 		0.50 ml 4.9e-4 0.001
Distilled water 1000.00 ml - -
1 Dissolve ingredients, adjust pH to 6.0 with 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 serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Adjust medium pH to 6.0, if necessary.
2 After inoculation, add by injection 3% (v/v) sterile air (usually 9 ml per bottle) and pressurize bottles with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.
561 SULFURIFERULA MEDIUM