Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
KH2PO4 | 0.50 | g | 0.498 | 3.66 | |
MgSO4 x 7 H2O | 0.40 | g | 0.398 | 1.616 | |
NaCl | 0.40 | g | 0.398 | 6.817 | |
NH4Cl | 0.40 | g | 0.398 | 7.448 | |
CaCl2 x 2 H2O | 0.05 | g | 0.05 | 0.339 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Selenite-tungstate solution | 1.00 | ml | - | - | |
FeSO4 x 7 H2O solution (0.1% w/v) | 2.00 | ml | - | - | |
Yeast extract
(OXOID) |
1.00 | g | 0.996 | - | |
Na-acetate | 1.00 | g | 0.996 | 12.142 | |
Na-formate | 2.00 | g | 1.992 | 29.292 | |
Sludge fluid | 50.00 | ml | - | - | |
Fatty acid mixture | 20.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
Methanol
(15% v/v) |
10.00 | ml | 7.888 | 246.191 | |
NaHCO3 | 1.00 | g | 0.996 | 11.856 | |
L-Cysteine HCl x H2O | 0.50 | g | 0.498 | 2.835 | |
Na2S x 9 H2O | 0.50 | g | 0.498 | 2.073 | |
Distilled water | 940.00 | ml | - | - | |
1 Dissolve ingredients except methanol, bicarbonate, cysteine and sulfide. Adjust pH of medium to 7.2 and sparge with 100% N2 gas for 30 - 45 min to make it anoxic. Then dispense medium under same gas atmosphere into anoxic Hungate-type tubes to 30% v/v of their volume and autoclave. Add methanol (from 15% v/v stock solution), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use check pH of complete medium and adjust to 7.6, if necessary. | |||||
2 After inoculation, add sterile 80% H2 and 20% CO2 gas mixture to 0.5 bar overpressure. After growth becomes visible overpressure of 80% H2 and 20% CO2 gas mixture can be increased to 1 bar. | |||||
3 Note: Use 10% (v/v) as inoculum. |
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] |
---|---|---|---|---|
NaOH | 0.5 | g | 0.5 | 12.501 |
Na2SeO3 x 5 H2O | 3.0 | mg | 0.003 | 0.011 |
Na2WO4 x 2 H2O | 4.0 | mg | 0.004 | 0.012 |
Distilled water | 1000.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Yeast extract | 4 | g | 4 | - | |
Sludge | 1000 | ml | - | - | |
1 Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours. Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at 8-12°C in the dark. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Isobutyric acid | 23 | ml | - | - | |
DL-2-Methylbutyric acid | 27 | ml | - | - | |
Valeric acid | 27 | ml | - | - | |
Isovaleric acid | 27 | ml | - | - | |
Distilled water | 896 | ml | - | - | |
1 Adjust pH to 7.5 with concentrated NaOH. |
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: | 11.01.24 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 7.6 |
Equipment needed: | Autoclave, Centrifuge, 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] |
---|---|---|
Methanol | 7.888 | 246.191 |
Na-formate | 1.992 | 29.292 |
Na-acetate | 0.996 | 12.142 |
NaHCO3 | 0.996 | 11.856 |
KH2PO4 | 0.498 | 3.66 |
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