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DSMZ

DSMZ 667 . DESULFALLAS THERMOSAPOVORANS MEDIUM

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Selected strain: Desulfallas thermosapovorans DSM 6562

This strain uses the standard medium without modifications.

Growth conditions: anaerobic 50 °C
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 973 ml
Solution B 30 ml
Solution C 1 ml
Solution D 5 ml
Solution E 1 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 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 is prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution D is autoclaved under 100% N2 gas. Solution E is prepared under 80% N2 and 20% CO2 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 at 7.0 - 7.2.

Solution A #

973
Compound Amount Unit
Na2SO4 3.00 g
KH2PO4 0.20 g
NH4Cl 1.00 g
NaCl 8.00 g
MgCl2 x 6 H2O 0.40 g
KCl 0.50 g
CaCl2 x 2 H2O 0.15 g
Trace element solution SL-10 1.50 ml
Selenite-tungstate solution 1.00 ml
Na-acetate 0.50 g
Na-butyrate 2.20 g
Yeast extract 0.40 g
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 970.00 ml

Solution B #

30
Compound Amount Unit
Na2CO3 1.5 g
Distilled water 30.0 ml

Solution C #

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

Solution D #

5
Compound Amount Unit
Na2S x 9 H2O 0.2 g
Distilled water 5.0 ml

Solution E #

1
Compound Amount Unit
Na-dithionite solution (5% w/v) 1 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.

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

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

Na-dithionite solution (5% w/v) #

10
Compound Amount Unit
NaOH
(1 N) 		1.0 ml
Na2S2O4 0.5 g
Distilled water 9.0 ml
1 Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.