DSMZ 855 . THAUERA AROMATICA MEDIUM (ANAEROB)

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Equipment needed: Autoclave Filter Gassing station Hungate tubes

Final gas composition: 80 % N2 20 % CO2 read more

Final pH: 7.2

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ml
Compound Amount Unit
Solution A 952 ml
Solution B 10 ml
Solution C 10 ml
Solution D 30 ml
Solution E 1 ml
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solutions B and C are autoclaved separately under 100% N2 gas. Solution D is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution E is prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions B to E are added to the sterile, cooled solution A in appropriate amounts in the sequence as indicated. Adjust pH of final medium to 7.2, if necessary.
2 For aerobic growth prepare medium under air. Omit carbonate and KNO3. 3,5-Dihydroxybenzoic acid or Na-benzoate as substrate can be replaced with 10 mM Na-malate. Adjust medium pH to 7.2.

Solution A #

952
Compound Amount Unit
Na2SO4 3.00 g
KH2PO4 0.20 g
NH4Cl 0.30 g
NaCl 1.00 g
MgCl2 x 6 H2O 0.40 g
KCl 0.50 g
CaCl2 x 2 H2O 0.15 g
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Distilled water 950.00 ml

Solution B #

10
Compound Amount Unit
3,5-Dihydroxybenzoic acid 0.6 g
Distilled water 10.0 ml

Solution C #

10
Compound Amount Unit
KNO3 0.6 g
Distilled water 10.0 ml

Solution D #

30
Compound Amount Unit
Na2CO3 1.5 g
Distilled water 30.0 ml

Solution E #

1
Compound Amount Unit
Wolin's vitamin solution (10x) 1 ml

Trace element solution SL-10 #

1000
Compound Amount Unit
HCl
(25%)
10.0 ml
FeCl2 x 4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2 x 4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2 x 6 H2O 190.0 mg
CuCl2 x 2 H2O 2.0 mg
NiCl2 x 6 H2O 24.0 mg
Na2MoO4 x 2 H2O 36.0 mg
Distilled water 990.0 ml
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Selenite-tungstate solution #

1000
Compound Amount Unit
NaOH 0.5 g
Na2SeO3 x 5 H2O 3.0 mg
Na2WO4 x 2 H2O 4.0 mg
Distilled water 1000.0 ml