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Main sol. 963

Internal Solution-ID: S1987

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.78 g 0.772 4.43
KH2PO4 0.75 g 0.742 5.451
Na3-EDTA 0.04 g 0.04 0.11
FeSO4 x 7 H2O 0.01 g 0.01 0.036
MgSO4 x 7 H2O 0.25 g 0.247 1.003
CaCl2 x 2 H2O 0.03 g 0.03 0.202
NaCl 0.20 g 0.198 3.385
NH4Cl 0.50 g 0.495 9.246
D-Glucose 0.50 g 0.495 2.745
Yeast extract
(OXOID)
0.50 g 0.495 -
Trace element solution 10.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
NaHCO3 5.00 g 4.946 58.871
Na2S2O3 x 5 H2O 1.24 g 1.227 4.942
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.25 g 0.247 1.408
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, thiosulfate, vitamins and cysteine), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add solid bicarbonate and adjust pH to 6.8 - 7.0. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add thiosulfate, vitamins and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of thiosulfate and vitamins should be sterilized by filtration. Adjust pH of complete medium to 6.5, if necessary.
963 THERMOANAEROMONAS MEDIUM