Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaCl | 20.00 | g | 19.782 | 338.508 | |
K2HPO4 | 0.09 | g | 0.089 | 0.511 | |
KH2PO4 | 0.07 | g | 0.069 | 0.509 | |
CaCl2 | 0.80 | g | 0.791 | 7.13 | |
NH4Cl | 0.25 | g | 0.247 | 4.623 | |
NaNO3 | 1.00 | g | 0.989 | 11.637 | |
MgSO4 x 7 H2O | 4.00 | g | 3.956 | 16.052 | |
MgCl2 x 6 H2O | 3.00 | g | 2.967 | 14.596 | |
KCl | 0.33 | g | 0.326 | 4.378 | |
Fe2(SO4)3 x H2O | 0.01 | g | 0.01 | - | |
Modified Wolin's mineral solution | 10.00 | ml | - | - | |
Na2S2O3 x 5 H2O | 1.00 | g | 0.989 | 3.985 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Sulfur
(powdered) |
10.00 | g | 9.891 | 308.425 | |
NaHCO3 | 1.00 | g | 0.989 | 11.774 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except sulfur, bicarbonate and vitamins), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.8 with NaOH and dispense under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (up to a volume of 20%) that contain already the appropriate amount of sulfur. Autoclave at a temperature of 110°C for 20 min. Add vitamins and bicarbonate to the autoclaved medium from sterile anoxic stock solutions. The solution of vitamins is sterilized by filtration and stored under 100% N2 gas, whereas the solution of bicarbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust the pH of the complete medium to 6.5. | |||||
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 | - | - |
Last modified: | 24.07.23 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Defined medium |
Final pH: | 6.8 |
Equipment needed: | Autoclave, 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] |
---|---|---|
Sulfur | 9.891 | 308.425 |
MgCl2 x 6 H2O | 2.967 | 14.596 |
NaHCO3 | 0.989 | 11.774 |
NaNO3 | 0.989 | 11.637 |
Na2S2O3 x 5 H2O | 0.989 | 3.985 |
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