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Main sol. 494

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Internal Solution-ID: S1005

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 150.00 g 148.368 2538.81
KCl 0.33 g 0.326 4.378
NH4Cl 0.33 g 0.326 6.102
KH2PO4 0.33 g 0.326 2.399
CaCl2 x 2 H2O 0.33 g 0.326 2.22
MgCl2 x 6 H2O 4.00 g 3.956 19.461
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 2.00 g 1.978 18.665
Yeast extract 0.05 g 0.049 -
Trimethylamine-HCl 2.40 g 2.374 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, yeast extract, trimethylamine, vitamins 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. Add yeast extract, trimethylamine, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Prior to use check pH of complete medium and adjust to 7.6 - 8.0 with a sterile anoxic stock solution of Na2CO3 (5% w/v), if necessary.
494 ACETOHALOBIUM MEDIUM