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Main sol. 135a

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Internal Solution-ID: S232

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 20.00 g 19.589 335.192
NH4Cl 1.00 g 0.979 18.311
KH2PO4 0.33 g 0.323 2.375
K2HPO4 0.45 g 0.441 2.53
MgSO4 x 7 H2O 0.10 g 0.098 0.397
Modified Wolin's mineral solution 20.00 ml - -
Yeast extract 2.00 g 1.959 -
Resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 5.00 g 4.897 58.295
Ethylene glycol 1.25 g 1.224 19.725
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.49 2.788
Na2S x 9 H2O 0.50 g 0.49 2.039
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, ethylene glycol, vitamins, cysteine and sulfide, bring to the boil and cool to room temperature under 80% N2 and 20% CO2 gas mixture. Add bicarbonate (solid) and equilibrate the medium with the gas until a pH of around 7.4 is reached. Then distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add ethylene glycol, vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 7.0 - 7.2, if necessary.
135a ACETOBACTERIUM MEDIUM (MARINE)