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Main sol. 525

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Internal Solution-ID: S1160

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.336 4.511
MgCl2 x 6 H2O 2.75 g 2.72 13.379
MgSO4 x 7 H2O 3.45 g 3.412 13.845
NH4Cl 0.25 g 0.247 4.623
K2HPO4 x 3 H2O 0.14 g 0.138 0.795
CaCl2 x 2 H2O 0.14 g 0.138 0.942
NaCl 100.00 g 98.912 1692.54
Modified Wolin's mineral solution 10.00 ml - -
Na-acetate 1.00 g 0.989 12.058
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 4.00 g 3.956 47.097
Na2CO3 0.50 g 0.495 4.666
Trimethylamine-HCl 5.00 g 4.946 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.495 2.816
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonates, trimethylamine, vitamins, cysteine and sulfide, then sparge medium with 80% N2 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 and autoclave. Complete medium by adding sterile anoxic stock solutions of trimethylamine, vitamins (sterilized by filtration), cysteine and sulfide prepared under 100% N2 gas atmosphere and bicarbonate and carbonate from sterile anoxic stock solutions prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 6.9 - 7.0, if necessary.
525 METHANOHALOPHILUS (MG) MEDIUM