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DSMZ

DSMZ 149 . MEGADESULFOVIBRIO MEDIUM

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Equipment needed: Autoclave Filter Gassing station Hungate tubes
Final gas composition: 80 % N2 20 % CO2 read more
Final pH: 7.0 - 7.2
Select a strain (optional):

ml
Compound Amount Unit
Solution A 970 ml
Solution B 20 ml
Solution C 1 ml
Solution D 10 ml
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution D is autoclaved separately under 100% N2 gas. To complete the medium appropriate amounts of solutions B to D are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.0 - 7.2.
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% inoculum.

Solution A #

970
Compound Amount Unit
KH2PO4 1.0 g
NH4Cl 0.5 g
MgSO4 x 7 H2O 0.4 g
Na2SO4 2.0 g
CaCl2 x 2 H2O 0.1 g
Trace element solution SL-6 1.0 ml
H2SO4
(2 N) 		1.0 ml
Na-L-lactate 2.0 g
Sodium resazurin
(0.1% w/v) 		0.5 ml
Distilled water 970.0 ml

Solution B #

20
Compound Amount Unit
Na2CO3 1 g
Distilled water 20 ml

Solution C #

1
Compound Amount Unit
Seven vitamins solution 1 ml

Solution D #

10
Compound Amount Unit
Na2S x 9 H2O 0.3 g
Distilled water 10.0 ml

Trace element solution SL-6 #

1000
Compound Amount Unit
ZnSO4 x 7 H2O 0.10 g
MnCl2 x 4 H2O 0.03 g
H3BO3 0.30 g
CoCl2 x 6 H2O 0.20 g
CuCl2 x 2 H2O 0.01 g
NiCl2 x 6 H2O 0.02 g
Na2MoO4 x 2 H2O 0.03 g
Distilled water 1000.00 ml

Seven vitamins solution #

1000
Compound Amount Unit
Vitamin B12 100 mg
p-Aminobenzoic acid 80 mg
D-(+)-biotin 20 mg
Nicotinic acid 200 mg
Calcium pantothenate 100 mg
Pyridoxine hydrochloride 300 mg
Thiamine-HCl x 2 H2O 200 mg
Distilled water 1000 ml