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Main sol. 898

Internal Solution-ID: S1848

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.2 1.467
MgCl2 x 6 H2O 0.10 g 0.1 0.491
KCl 0.20 g 0.2 2.677
NH4Cl 1.00 g 0.998 18.658
NaCl 5.00 g 4.99 85.387
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
Na2CO3 2.76 g 2.754 25.989
NaHCO3 10.00 g 9.98 118.8
Yeast extract 0.50 g 0.499 -
D-Glucose 5.00 g 4.99 27.698
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.499 2.078
Distilled water 1000.00 ml - -
1 Dissolve all ingredients except carbonates, yeast extract, glucose, vitamins and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add carbonate and hydrogencarbonate, adjust pH to 9.0 - 9.5, dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add yeast extract, glucose, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 9.5 - 9.7.
898 ANOXYBACILLUS MEDIUM