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DSMZ

DSMZ 875 . DESULFACINUM HYDROTHERMALE 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 - 7.3
Select a strain (optional):

ml
Compound Amount Unit
Solution A 950 ml
Solution B 1 ml
Solution C 30 ml
Solution D 1 ml
Solution E 10 ml
Solution F 10 ml
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 - 45 min), then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solutions B, E and F are autoclaved separately under 100% N2 gas atmosphere. Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution D is prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.0 - 7.3.

Solution A #

950
Compound Amount Unit
NaCl 26.00 g
MgCl2 x 6 H2O 5.60 g
CaCl2 x 2 H2O 1.40 g
MgSO4 x 7 H2O 6.80 g
NH4Cl 0.25 g
KH2PO4 0.20 g
KCl 0.72 g
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 950.00 ml

Solution B #

1
Compound Amount Unit
Trace element solution SL-10 1 ml

Solution C #

30
Compound Amount Unit
Na2CO3 1.5 g
Distilled water 30.0 ml

Solution D #

1
Compound Amount Unit
Seven vitamins solution 1 ml

Solution E #

10
Compound Amount Unit
Na-DL-lactate 2.5 g
Distilled water 10.0 ml

Solution F #

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

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