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Main sol. 876

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Internal Solution-ID: S1815

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 4.00 g 3.98 28.021
KH2PO4 0.20 g 0.199 1.462
NH4Cl 0.25 g 0.249 4.651
NaCl 20.00 g 19.901 340.529
MgCl2 x 6 H2O 9.80 g 9.751 47.964
KCl 0.50 g 0.498 6.674
CaCl2 x 2 H2O 0.10 g 0.1 0.677
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 2.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2CO3 1.00 g 0.995 9.388
Na-caproate 0.50 g 0.498 3.602
Na-butyrate 0.30 g 0.299 2.712
Wolin's vitamin solution (10x) 1.00 ml - -
Seven vitamins solution 1.00 ml - -
Na2S x 9 H2O 0.40 g 0.398 1.657
Distilled water 1000.00 ml - -
1 Dissolve all ingredients except carbonate, caproate, butyrate, vitamins and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add caproate, butyrate, vitamins 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 atmosphere. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 6.7 - 6.9, if necessary.
2 Note: Growth can be stimulated by supplementing the medium with 1% (v/v) of fatty acid mixture (see medium 119).
876 DESULFOTHERMUS NAPHTHAE MEDIUM