Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaCl | 13.85 | g | 13.512 | 231.215 | |
MgSO4 x 7 H2O | 3.50 | g | 3.415 | 13.854 | |
MgCl2 x 6 H2O | 2.75 | g | 2.683 | 13.197 | |
KCl | 0.33 | g | 0.322 | 4.319 | |
NaBr | 0.05 | g | 0.049 | 0.474 | |
H3BO3 | 15.00 | mg | 0.015 | 0.237 | |
SrCl2 x 6 H2O
(0.1% w/v) |
7.00 | ml | 0.007 | 0.026 | |
(NH4)2SO4 | 10.00 | mg | 0.01 | 0.074 | |
Citric acid
(0.1% w/v) |
5.00 | ml | 0.005 | 0.025 | |
KI
(0.01% w/v) |
0.50 | ml | 4.9e-5 | 1.9e-4 | |
CaCl2 x 2 H2O | 0.75 | g | 0.732 | 4.977 | |
KH2PO4 | 0.50 | g | 0.488 | 3.585 | |
NiCl2 x 6 H2O
(0.1% w/v) |
2.00 | ml | 0.002 | 0.015 | |
Modified Wolin's mineral solution | 10.00 | ml | - | - | |
Sulfur
(powdered) |
30.00 | g | 29.268 | 912.638 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Yeast extract
(OXOID) |
2.00 | g | 1.951 | - | |
Na2S x 9 H2O | 0.50 | g | 0.488 | 2.031 | |
Distilled water | 1000.00 | ml | - | - | |
1 Prepare the medium without sulfur, yeast extract and sodium sulfide, adjust pH to 5.0 - 5.5 with 10 N sulfuric acid, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under 80% H2 and CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials that contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. For sterilization of medium heat vessels for at least 1 hour to 90 - 100°C on each of 3 successive days. Do not autoclave! Complete the medium by adding yeast extract and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 5.5, if necessary. | |||||
2 After inoculation pressurize the 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. |
Last modified: | 11.08.23 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 5.5 |
Equipment needed: | Gassing station, 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 .
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Sulfur | 29.268 | 912.638 |
MgCl2 x 6 H2O | 2.683 | 13.197 |
Yeast extract | 1.951 | - |
KH2PO4 | 0.488 | 3.585 |
Na2S x 9 H2O | 0.488 | 2.031 |
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