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Main sol. 870

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Internal Solution-ID: S1798

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.14 g 0.138 1.017
MgCl2 x 6 H2O 0.20 g 0.198 0.972
CaCl2 x 2 H2O 0.15 g 0.148 1.008
NH4Cl 0.54 g 0.534 9.976
Modified Wolin's mineral solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 1.00 g 0.988 9.323
Na-crotonate solution (1 M) 10.00 ml - -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.30 g 0.296 1.234
L-Cysteine HCl x H2O 0.30 g 0.296 1.688
Distilled water 1000.00 ml - -
1 Dissolve all ingredients except carbonate, crotonate, vitamins, sulfide and cysteine, 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 crotonate, vitamins, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Sterilize vitamins by filtration. Adjust pH of complete medium to 7.0, if necessary.
870 SYNTROPHOTHERMUS MEDIUM