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Main sol. 135b

Internal Solution-ID: S233

Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
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 -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Na2CO3 1.50 g 1.469 13.862
Na-syringate 1.00 g 0.979 4.449
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.49 2.788
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, syringate, vitamins and cysteine, bring to the boil and cool to room temperature under 80% N2 and 20% CO2 gas atmosphere. Thereafter, dispense under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Add syringate, vitamins and cysteine 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. Vitamins should be sterilized by filtration. The pH of the completed medium should be at 7.4.
2 Note: After growth has been started add an additional amount of syringate to reach a final concentration of 2.00 g/l.
135b ACETOBACTERIUM DEHALOGENANS MEDIUM