Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
(NH4)2SO4 | 1.30 | g | 1.286 | 9.731 | |
KH2PO4 | 0.28 | g | 0.277 | 2.035 | |
MgSO4 x 7 H2O | 0.25 | g | 0.247 | 1.003 | |
CaCl2 x 2 H2O | 0.07 | g | 0.069 | 0.471 | |
FeCl3 x 6 H2O | 0.02 | g | 0.02 | 0.073 | |
Allen's trace element solution | 10.00 | ml | - | - | |
Trisodium citrate x 2 H2O | 2.94 | g | 2.908 | 9.888 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Sulfur
(powdered) |
10.00 | g | 9.891 | 308.425 | |
Yeast extract
(OXOID) |
0.50 | g | 0.495 | - | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Na2S x 9 H2O | 0.50 | g | 0.495 | 2.059 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except sulfur, yeast extract, vitamins and sulfide), adjust pH to 3.5 with 10 N H2SO4 and 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 (e.g., 20 ml medium in 100 ml bottles) containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 4.0 - 4.5, if necessary. | |||||
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
MnCl2 x 4 H2O | 180 | mg | 0.18 | 0.91 | |
Na2B4O7 x 10 H2O | 450 | mg | 0.45 | - | |
ZnSO4 x 7 H2O | 22 | mg | 0.022 | 0.077 | |
CuCl2 x 2 H2O | 5 | mg | 0.005 | 0.029 | |
Na2MoO4 x 2 H2O | 3 | mg | 0.003 | 0.012 | |
VOSO4 x 2 H2O | 3 | mg | 0.003 | 0.015 | |
CoSO4 x 7 H2O | 1 | mg | 1.0e-3 | 0.006 | |
Distilled water | 1000 | ml | - | - | |
1 Adjust pH of final solution to 2 with 1 N HCl. |
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: | 17.10.22 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 4.0 - 4.5 |
Equipment needed: | Autoclave, Filter, Gassing station, Hungate tubes, Serum bottle, Water bath |
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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 |
Trisodium citrate x 2 H2O | 2.908 | 9.888 |
(NH4)2SO4 | 1.286 | 9.731 |
Na2S x 9 H2O | 0.495 | 2.059 |
Yeast extract | 0.495 | - |
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