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Main sol. 819

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Internal Solution-ID: S1667

Equipment needed: Autoclave Hungate tubes 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 0.2 g 0.2 0.984
CaCl2 x 2 H2O 0.1 g 0.1 0.68
KCl 0.2 g 0.2 2.683
NaCl 50.0 g 50 855.578
Yeast extract 5.0 g 5 -
Modified Wolin's mineral solution 10.0 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 -
L-Cysteine HCl x H2O 0.3 g 0.3 1.708
Na2CO3 1.0 g 1 9.435
Na2S x 9 H2O 0.3 g 0.3 1.249
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except cysteine, carbonate and sulfide), sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then, add cysteine, distribute medium under 80% N2 and 20% CO2 gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Check pH of complete medium and adjust to pH 7.0, if necessary.
819 SEDIMINISPIROCHAETA SMARAGDINAE MEDIUM