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Main sol. 786

Internal Solution-ID: S1592

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 1 18.695
K2HPO4 0.3 g 0.3 1.722
KH2PO4 0.3 g 0.3 2.205
MgCl2 x 6 H2O 3.0 g 3 14.756
CaCl2 x 2 H2O 0.1 g 0.1 0.68
KCl 0.1 g 0.1 1.341
Na-acetate 0.5 g 0.5 6.095
Yeast extract
(OXOID)
1.0 g 1 -
Trypticase peptone
(BD BBL)
5.0 g 5 -
NaCl 30.0 g 30 513.347
Modified Wolin's mineral solution 10.0 ml - -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 -
Na2CO3 1.5 g 1.5 14.153
Na2S2O3 x 5 H2O 5.0 g 5 20.146
L-Cysteine HCl x H2O 0.5 g 0.5 2.847
Na2S x 9 H2O 0.5 g 0.5 2.082
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, thiosulfate, 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 thiosulfate, cysteine 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. Stock solutions of thiosulfate and vitamins should be sterilized by filtration. The pH of the complete medium should be 7.2 - 7.4.
786 DETHIOSULFOVIBRIO PEPTIDOVORANS MEDIUM