Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
K2HPO4 | 0.35 | g | 0.342 | 1.962 | |
KH2PO4 | 0.23 | g | 0.225 | 1.651 | |
NH4Cl | 0.50 | g | 0.488 | 9.128 | |
MgSO4 x 7 H2O | 0.50 | g | 0.488 | 1.981 | |
CaCl2 x 2 H2O | 0.25 | g | 0.244 | 1.661 | |
NaCl | 2.25 | g | 2.197 | 37.599 | |
FeSO4 x 7 H2O solution (0.1% w/v) | 2.00 | ml | - | - | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Yeast extract
(OXOID) |
2.00 | g | 1.953 | - | |
Casitone
(BD BBL) |
2.00 | g | 1.953 | - | |
Na-acetate | 2.50 | g | 2.441 | 29.761 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
NaHCO3 | 2.50 | g | 2.441 | 29.062 | |
2-Mercaptoethanesulfonate
(coenzyme M) |
0.14 | g | 0.137 | 0.961 | |
Methanol
(50% v/v) |
20.00 | ml | 15.469 | 482.765 | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
L-Cysteine HCl x H2O | 0.36 | g | 0.352 | 2.002 | |
Na2S x 9 H2O | 0.36 | g | 0.352 | 1.464 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.5 and dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N2 gas atmosphere as concentrated solutions in tightly closed tubes. Vitamins and coenzyme M are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 7.0 - 7.2, if necessary. | |||||
2 After inoculation, pressurize culture vessels with sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure. |
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. |
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] | |
---|---|---|---|---|---|
FeSO4 x 7 H2O | 1 | g | 1 | 3.597 | |
H2SO4
(0.1 N) |
1000 | ml | - | - | |
1 The ferrous sulfate solution is not stable and should be freshly prepared. |
Last modified: | 01.12.22 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 7.0 - 7.2 |
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 .
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Methanol | 15.469 | 482.765 |
Na-acetate | 2.441 | 29.761 |
NaHCO3 | 2.441 | 29.062 |
NaCl | 2.197 | 37.599 |
Yeast extract | 1.953 | - |
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