MediaDive DSMZ
Content
Media Solutions Ingredients Strains Steps Gas
Search
Medium finder Solution finder Taxonomy search Isolation sources
Tools
Medium builder NEW Unit converter Compare media Compare solutions Prediction SOON
DSMZ

DSMZ 346 . DESULFALLAS MEDIUM

What is this page?
Equipment needed: Autoclave Centrifuge Filter Gassing station Hungate tubes
Final gas composition: 100 % N2 read more
Final pH: 7.2 - 7.5
Select a strain (optional):

ml
Compound Amount Unit
Solution A 966.0 ml
Solution B 20.0 ml
Solution C 11.0 ml
Solution D 10.0 ml
Solution E 1.0 ml
Solution F 1.0 ml
Solution G 1.0 ml
Solution H 1.7 ml
1 Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic. Thereafter, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Anoxic stock solutions C, D, E, F, and H are prepared under 100% N2 gas atmosphere and solution G under 80% N2 and 20% CO2 gas atmosphere. Filter sterilize solutions D, F and G. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 7.2 - 7.5, if necessary.

Solution A #

966
Compound Amount Unit
Na2SO4 3.00 g
KH2PO4 0.20 g
NH4Cl 0.30 g
CaCl2 x 2 H2O 0.15 g
MgCl2 x 6 H2O 0.40 g
NaCl 1.00 g
Trace element solution SL-10 1.00 ml
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 965.00 ml

Solution B #

20
Compound Amount Unit
NaHCO3 1 g
Distilled water 20 ml

Solution C #

11
Compound Amount Unit
Ethanol 1 ml
Distilled water 10 ml

Solution D #

10
Compound Amount Unit
Na-benzoate 0.7 g
Distilled water 10.0 ml

Solution E #

1
Compound Amount Unit
Clarified rumen fluid 1 ml

Solution F #

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

Solution G #

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

Solution H #

2
Compound Amount Unit
Na2S x 9 H2O 50.0 mg
Distilled water 1.7 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.

Clarified rumen fluid #

1000
1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.

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) #

1000
Compound Amount Unit
NaHCO3 50 g
Na2S2O4 50 g
Distilled water 1000 ml
1 Dissolve NaHCO3 in water and make the solution anoxic by sparging with 80% N2 and 20% CO2 gas mixture. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in the dark and refrigerated. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.