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Main sol. 713

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Internal Solution-ID: S1464

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.5 g 0.5 9.348
NaCl 60.0 g 60 1026.69
MgCl2 x 6 H2O 2.4 g 2.4 11.805
KCl 1.0 g 1 13.414
KH2PO4 0.4 g 0.4 2.939
Na2-EDTA 0.5 g 0.5 1.487
Yeast extract 2.0 g 2 -
Modified Wolin's mineral solution 10.0 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 -
Na2CO3 1.0 g 1 9.435
Trimethylamine-HCl 10.0 g 10 -
Wolin's vitamin solution 5.0 ml - -
CaCl2 x 2 H2O 2.0 g 2 13.604
L-Cysteine HCl x H2O 0.5 g 0.5 2.847
Na2S x 9 H2O 0.5 g 0.5 2.082
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except carbonate, trimethylamine, vitamins, calcium chloride, cysteine and sulfide), bring medium to the boil, then cool to room temperature under 80% N2 and 20% CO2 gas mixture. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add trimethylamine, calcium chloride, cysteine 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 mixture. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.8 - 7.0, if necessary.
713 METHANOHALOPHILUS EUHALOBIUS MEDIUM