Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Supplement medium with 1.00 g/l NaNO3 and omit pyruvate, lactate, yeast extract and dithionite. Upon autoclaving add 10.00 g/l sterile sulfur powder (sterilized by steaming for 3 hours on each of 3 successive days) and adjust pH to 6.0 - 6.5. After inoculation do not pressurize with 80% H2 and 20% CO2 gas mixture and add sterile air in an amount that is equivalent to a volume of 10% of the headspace.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaCl | 30.00 | g | 29.644 | 507.26 | |
K2HPO4 | 0.14 | g | 0.138 | 0.794 | |
CaCl2 x 2 H2O | 0.14 | g | 0.138 | 0.941 | |
MgSO4 x 7 H2O | 3.40 | g | 3.36 | 13.631 | |
MgCl2 x 6 H2O | 4.18 | g | 4.13 | 20.317 | |
KCl | 0.33 | g | 0.326 | 4.374 | |
NH4Cl | 0.25 | g | 0.247 | 4.618 | |
Fe(NH4)2(SO4)2 x 6 H2O | 0.01 | g | 0.01 | - | |
Modified Wolin's mineral solution | 10.00 | ml | - | - | |
Na2CO3 | 1.50 | g | 1.482 | 13.985 | |
Na-pyruvate* | 0.50 | g | 0.494 | 4.49 | |
Na-lactate* | 0.50 | g | 0.494 | 4.409 | |
Yeast extract* | 0.10 | g | 0.099 | - | |
Na2S2O3 x 5 H2O | 1.50 | g | 1.482 | 5.972 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Na-dithionite solution (5% w/v)* | 1.00 | ml | - | - | |
Sulfur powder* | 10.00 | g | - | - | |
NaNO3* | 1.00 | g | - | - | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except carbonate, pyruvate, lactate, yeast extract, thiosulfate, vitamins and dithionite), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials up to a volume of 20% and autoclave. Add pyruvate, lactate, yeast extract, thiosulfate and vitamins to the autoclaved medium from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate and dithionite from sterile anoxic stock solutions prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins, thiosulfate and dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.7. | |||||
2 After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Nitrilotriacetic acid | 1.50 | g | 1.5 | 7.848 | |
MgSO4 x 7 H2O | 3.00 | g | 3 | 12.172 | |
MnSO4 x H2O | 0.50 | g | 0.5 | 2.958 | |
NaCl | 1.00 | g | 1 | 17.112 | |
FeSO4 x 7 H2O | 0.10 | g | 0.1 | 0.36 | |
CoSO4 x 7 H2O | 0.18 | g | 0.18 | 1.161 | |
CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.68 | |
ZnSO4 x 7 H2O | 0.18 | g | 0.18 | 0.626 | |
CuSO4 x 5 H2O | 0.01 | g | 0.01 | 0.04 | |
AlK(SO4)2 x 12 H2O | 0.02 | g | 0.02 | 0.042 | |
H3BO3 | 0.01 | g | 0.01 | 0.162 | |
Na2MoO4 x 2 H2O | 0.01 | g | 0.01 | 0.041 | |
NiCl2 x 6 H2O | 0.03 | g | 0.03 | 0.231 | |
Na2SeO3 x 5 H2O | 0.30 | mg | 3.0e-4 | 0.001 | |
Na2WO4 x 2 H2O | 0.40 | mg | 4.0e-4 | 0.001 | |
Distilled water | 1000.00 | ml | - | - | |
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. |
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: | 17.10.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | * 6.0 - 6.5 |
Equipment needed: | Autoclave, Filter, Gassing station, Hungate tubes |
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Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Sulfur powder | 9.881 | 38.524 |
MgCl2 x 6 H2O | 4.13 | 20.317 |
Na2CO3 | 1.482 | 13.985 |
Na2S2O3 x 5 H2O | 1.482 | 5.972 |
NaNO3 | 0.988 | 11.626 |
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