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Main sol. 853a

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Internal Solution-ID: S1743

Equipment needed: Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.99 18.51
K2HPO4 0.30 g 0.297 1.705
KH2PO4 0.30 g 0.297 2.183
MgCl2 x 6 H2O 3.00 g 2.97 14.61
CaCl2 x 2 H2O 0.10 g 0.099 0.673
KCl 1.00 g 0.99 13.281
NaCl 70.00 g 69.307 1185.95
Yeast extract
(OXOID) 		1.00 g 0.99 -
Trypticase peptone
(BD BBL) 		1.00 g 0.99 -
Na-acetate x 3 H2O 0.50 g 0.495 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
Na2S2O3 x 5 H2O 3.16 g 3.129 12.606
L-Cysteine HCl x H2O 0.50 g 0.495 2.819
Na2CO3 1.50 g 1.485 14.012
D-Glucose 3.60 g 3.564 19.784
Na2S x 9 H2O 0.30 g 0.297 1.237
Distilled water 1000.00 ml - -
1 Dissolve ingredients, except thiosulfate, cysteine, carbonate, glucose, and sulfide, then sparge medium for 30 - 45 min with 80% N2 and 20% CO2 gas mixture to make it anoxic. Add thiosulfate and cysteine, then adjust pH to 7.0 and dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. After autoclaving, add glucose 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. Adjust the pH of the complete medium to 7.2 - 7.4, if necessary.
853a FUSIBACTER TUNISIENSIS MEDIUM