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Main sol. 1211

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Internal Solution-ID: S2415

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.05 g 0.05 0.286
KH2PO4 0.09 g 0.09 0.659
NH4Cl 0.25 g 0.249 4.66
MgSO4 x 7 H2O 0.25 g 0.249 1.011
CaCl2 x 2 H2O 0.15 g 0.15 1.017
Yeast extract 5.00 g 4.985 -
Trace element solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 0.25 g 0.249 2.352
Na2S2O3 x 5 H2O 2.50 g 2.493 10.043
Wolin's vitamin solution 2.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.249 1.419
Na2S x 9 H2O 0.25 g 0.249 1.038
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, thiosulfate, vitamins and reducing agents) and 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 thiosulfate, vitamin solution, 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 mixture. The pH of the complete medium should be at 6.5 - 6.8.
1211 CALDISERICUM MEDIUM