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Main sol. 1477

Internal Solution-ID: S3025

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.3 g 0.299 1.719
KH2PO4 0.3 g 0.299 2.2
NH4Cl 1.0 g 0.998 18.658
MgCl2 x 6 H2O 1.5 g 1.497 7.363
CaCl2 x 2 H2O 0.1 g 0.1 0.679
NaCl 20.0 g 19.96 341.548
KCl 0.1 g 0.1 1.339
Trace element solution 1.0 ml - -
Yeast extract 1.0 g 0.998 -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 0.002
NaHCO3 0.5 g 0.499 5.94
D-Glucose 3.6 g 3.593 19.942
Wolin's vitamin solution (10x) 1.0 ml - -
Distilled water 1000.0 ml - -
1 Dissolve ingredients except bicarbonate, glucose and vitamins, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose and vitamins (sterilized by filtration) from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.0 - 7.2, if necessary.
1477 CALORAMATOR QUIMBAYENSIS MEDIUM