Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
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. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2SO4 | 3.00 | g | 3.106 | 21.864 |
KH2PO4 | 0.20 | g | 0.207 | 1.521 |
NH4Cl | 0.30 | g | 0.311 | 5.806 |
CaCl2 x 2 H2O | 0.15 | g | 0.155 | 1.056 |
MgCl2 x 6 H2O | 0.40 | g | 0.414 | 2.037 |
NaCl | 1.00 | g | 1.035 | 17.714 |
Trace element solution SL-10 | 1.00 | ml | - | - |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.2e-4 | 0.002 |
Distilled water | 965.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
NaHCO3 | 1 | g | 50 | 595.188 |
Distilled water | 20 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na-benzoate | 0.7 | g | 70 | 485.764 |
Distilled water | 10.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2S x 9 H2O | 50.0 | mg | 25 | 104.088 |
Distilled water | 1.7 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
HCl
(25%) |
10.0 | ml | 2.5 | 68.568 | |
FeCl2 x 4 H2O | 1.5 | g | 1.5 | 7.545 | |
ZnCl2 | 70.0 | mg | 0.07 | 0.514 | |
MnCl2 x 4 H2O | 100.0 | mg | 0.1 | 0.505 | |
H3BO3 | 6.0 | mg | 0.006 | 0.097 | |
CoCl2 x 6 H2O | 190.0 | mg | 0.19 | 1.463 | |
CuCl2 x 2 H2O | 2.0 | mg | 0.002 | 0.012 | |
NiCl2 x 6 H2O | 24.0 | mg | 0.024 | 0.185 | |
Na2MoO4 x 2 H2O | 36.0 | mg | 0.036 | 0.149 | |
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. |
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. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Biotin | 20 | mg | 0.02 | 0.082 |
Folic acid | 20 | mg | 0.02 | 0.045 |
Pyridoxine hydrochloride | 100 | mg | 0.1 | 0.486 |
Thiamine HCl | 50 | mg | 0.05 | 0.148 |
Riboflavin | 50 | mg | 0.05 | 0.133 |
Nicotinic acid | 50 | mg | 0.05 | 0.406 |
Calcium D-(+)-pantothenate | 50 | mg | 0.05 | 0.21 |
Vitamin B12 | 1 | mg | 1.0e-3 | 7.4e-4 |
p-Aminobenzoic acid | 50 | mg | 0.05 | 0.365 |
(DL)-alpha-Lipoic acid | 50 | mg | 0.05 | 0.242 |
Distilled water | 1000 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaHCO3 | 50 | g | 50 | 595.188 | |
Na2S2O4 | 50 | g | 50 | 287.175 | |
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. |
Last modified: | 11.10.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 7.2 - 7.5 |
Equipment needed: | Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes |
Edit in Medium builder |
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
All strains for this medium: | 1 |
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Na2SO4 | 2.964 | 20.87 |
NaCl | 0.988 | 16.909 |
Na-benzoate | 0.692 | 4.8 |
MgCl2 x 6 H2O | 0.395 | 1.944 |
NH4Cl | 0.296 | 5.542 |
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