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Main sol. 481

Internal Solution-ID: S983

Equipment needed: Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.4 g 0.396 2.274
NH4Cl 1.0 g 0.99 18.51
Yeast extract 2.0 g 1.98 -
Trypticase 2.0 g 1.98 -
MgCl2 x 6 H2O 1.0 g 0.99 4.87
CaCl2 x 2 H2O 0.4 g 0.396 2.694
Modified Wolin's mineral solution 10.0 ml - -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
Na2CO3 1.5 g 1.485 14.012
Na2S x 9 H2O 0.3 g 0.297 1.237
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate 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 carbonate after autoclaving from an anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmospher and sulfide from a sterile anoxic stock solution autoclaved under 100% N2 gas. Adjust pH of the complete medium to pH 7.0.
481 COPROTHERMOBACTER PROTEOLYTICUS MEDIUM