Main sol. 135c

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