DSMZ 193 . DESULFOBACTER MEDIUM

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Equipment needed: Autoclave Filter Gassing station Hungate tubes

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

Final pH: 7.1 - 7.4

Select a strain (optional):

ml
Compound Amount Unit
Solution A 942 ml
Solution B 30 ml
Solution C 20 ml
Solution D 1 ml
Solution E 10 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 in anoxic Hungate-type tubes or serum vials and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and E are autoclaved under 100% N2 gas. Solution D is prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.1 - 7.4.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the beginning. For transfers use 5 - 10% (v/v) inoculum.

Solution A #

942
Compound Amount Unit
Na2SO4 3.00 g
KH2PO4 0.20 g
NH4Cl 0.30 g
NaCl 7.00 g
MgCl2 x 6 H2O 1.30 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
Sodium resazurin
(0.1% w/v)
0.50 ml
Distilled water 940.00 ml

Solution B #

30
Compound Amount Unit
Na2CO3 1.5 g
Distilled water 30.0 ml

Solution C #

20
Compound Amount Unit
Na-acetate x 3 H2O 2.5 g
Distilled water 20.0 ml

Solution D #

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

Solution E #

10
Compound Amount Unit
Na2S x 9 H2O 0.4 g
Distilled water 10.0 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

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.