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DSMZ

DSMZ 509 . SPIROCHAETA THERMOPHILA MEDIUM

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Equipment needed: Autoclave Filter Gassing station Hungate tubes
Final gas composition: 80 % N2 20 % CO2 read more
Final pH: 7.2 - 7.4
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ml
Compound Amount Unit
Solution A 952 ml
Solution B 30 ml
Solution C 1 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. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C, E and F are autoclaved under 100% N2 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 7.2 - 7.4.

Solution A #

952
Compound Amount Unit
NaCl 15.00 g
MgCl2 x 6 H2O 2.30 g
KCl 0.50 g
NH4Cl 0.30 g
KH2PO4 0.20 g
CaCl2 x 2 H2O 0.03 g
Selenite-tungstate solution 1.00 ml
Yeast extract 1.00 g
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 950.00 ml

Solution B #

30
Compound Amount Unit
Na2CO3 1.5 g
Distilled water 30.0 ml

Solution C #

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

Solution D #

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

Solution E #

10
Compound Amount Unit
Starch 1 g
Distilled water 10 ml

Solution F #

10
Compound Amount Unit
Na2S x 9 H2O 0.3 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.

Wolin's vitamin solution (10x) #

1000
Compound Amount Unit
Biotin 20 mg
Folic acid 20 mg
Pyridoxine hydrochloride 100 mg
Thiamine HCl 50 mg
Riboflavin 50 mg
Nicotinic acid 50 mg
Calcium D-(+)-pantothenate 50 mg
Vitamin B12 1 mg
p-Aminobenzoic acid 50 mg
(DL)-alpha-Lipoic acid 50 mg
Distilled water 1000 ml