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DSMZ

DSMZ 194a . DESULFOTOMACULUM OX39 MEDIUM (XYLENE)

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Equipment needed: Autoclave Filter Gassing station Serum bottle
Final gas composition: 80 % N2 20 % CO2 read more
Final pH: 7.2 - 7.4
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ml
Compound Amount Unit
Solution A 932 ml
Solution B 1 ml
Solution C 30 ml
Solution D 20 ml
Solution E 1 ml
Solution F 10 ml
Solution G 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 serum vials (e.g., 50 ml medium in 100 ml serum bottles) and autoclaved. Solutions B, D and G are autoclaved separately under 100% N2 gas. Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solutions E and F are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions B to G are added to the sterile, cooled solution A in appropriate amounts in the sequence as indicated. Final pH of the medium should be 7.2 - 7.4.
2 Note: For transfers use 5 - 10% (v/v) inoculum. Incubate tubes in a slanted position.

Solution A #

932
Compound Amount Unit
Na2SO4 1.40 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
Selenite-tungstate solution 1.00 ml
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 930.00 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 #

20
Compound Amount Unit
m-Xylene 0.3 ml
2,2,4,4,6,8,8-Heptamethylnonane 20.0 ml

Solution E #

1
Compound Amount Unit
Seven vitamins solution 1 ml

Solution F #

10
Compound Amount Unit
FeSO4 x 7 H2O 0.8 g
H2SO4
(0.2 N) 		10.0 ml

Solution G #

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.

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