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Main sol. 135c

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Internal Solution-ID: S234

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.979 18.311
KH2PO4 0.33 g 0.323 2.375
K2HPO4 0.45 g 0.441 2.53
MgSO4 x 7 H2O 0.10 g 0.098 0.397
Modified Wolin's mineral solution 20.00 ml - -
Yeast extract 2.00 g 1.959 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.00 g 0.979 11.659
D-Fructose
optional 		10.00 g 9.794 54.365
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.49 2.788
Na2S x 9 H2O 0.50 g 0.49 2.039
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, fructose, vitamins, cysteine and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add fructose, vitamins, cysteine and sulfide 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. The pH of the complete medium should be at 6.5.
2 Note: For autotrophic growth fructose is omitted and a gas atmosphere of 80% H2 and 20% CO2 is used.
135c MOORELLA THERMOACETICA MEDIUM