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Main sol. 794

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Internal Solution-ID: S1621

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.99 18.51
K2HPO4 0.3 g 0.297 1.705
KH2PO4 0.3 g 0.297 2.183
MgCl2 x 6 H2O 0.2 g 0.198 0.974
CaCl2 x 2 H2O 0.1 g 0.099 0.673
KCl 0.1 g 0.099 1.328
Na-acetate 0.5 g 0.495 6.035
NaCl 10.0 g 9.901 169.421
Yeast extract
(BD Bacto) 		2.0 g 1.98 -
Trypticase peptone
(BD BBL) 		2.0 g 1.98 -
Modified Wolin's mineral solution 10.0 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
L-Cysteine HCl x H2O 0.5 g 0.495 2.819
NaHCO3 2.0 g 1.98 23.572
D-Xylose 3.0 g 2.97 19.785
Na2S2O3 x 5 H2O 5.0 g 4.951 19.947
Na2S x 9 H2O 0.5 g 0.495 2.061
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except cysteine, bicarbonate, xylose, thiosulfate and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve cysteine, adjust pH to 6.5, then dispense medium under 80% N2 and 20% CO2 gas mixture into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. After sterilization add thiosulfate 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. Xylose is prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0 - 7.2, if necessary.
794 PSEUDOTHERMOTOGA HYPOGEA MEDIUM