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Main sol. 664

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Internal Solution-ID: S1396

Equipment needed: 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
NaCl 10.0 g 9.901 169.421
Modified Wolin's mineral solution 10.0 ml - -
Na-acetate 0.5 g 0.495 6.035
Yeast extract
(OXOID) 		5.0 g 4.951 -
Trypticase peptone
(BD BBL) 		5.0 g 4.951 -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
Na2CO3 1.5 g 1.485 14.012
Na2S2O3 x 5 H2O 5.0 g 4.951 19.947
D-Glucose 4.0 g 3.96 21.983
L-Cysteine HCl x H2O 0.5 g 0.495 2.819
Na2S x 9 H2O 0.5 g 0.495 2.061
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, thiosulfate, glucose 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, glucose, 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. The pH of the complete medium should be adjusted to 7.5, if necessary.
664 PSEUDOTHERMOTOGA ELFII MEDIUM