Main sol. 298

Equipment needed: Autoclave Hungate tubes Filter Gassing station

Compound	Amount	Unit	Conc. [g/L]	Conc. [mM]
KH₂PO₄	0.20	g	0.2	1.47
NH₄Cl	0.25	g	0.25	4.674
NaCl	1.00	g	1	17.112
MgCl₂ x 6 H₂O	0.40	g	0.4	1.968
KCl	0.50	g	0.5	6.707
CaCl₂ x 2 H₂O	0.15	g	0.15	1.02
Trace element solution SL-10	1.00	ml	-	-
Sodium resazurin (0.1% w/v)	0.50	ml	5.0e-4	-
Na₂CO₃	1.50	g	1.5	14.153
2,3-butanediol	0.90	g	0.9	9.987
Na₂S x 9 H₂O	0.36	g	0.36	1.499
Distilled water	1000.00	ml	-	-
1 Dissolve ingredients except carbonate, 2,3-butanediol and sulfide, then sparge medium with 80% N₂ and 20% CO₂ gas mixture for 30 - 45 min to make it anoxic. Dispense medium in anoxic Hungate-type tubes or serum vials under the same gas atmosphere and autoclave. Add 2,3-butanediol and sulfide 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. Prior to use adjust pH of complete medium to 7.2 - 7.5.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution freshly prepared under N₂ and filter-sterilized) may stimulate growth of some strains at the beginning.

298	PELOBACTER PROPIONICUS MEDIUM (FRESHWATER)