Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NH4Cl | 0.33 | g | 0.324 | 6.066 | |
KCl | 0.33 | g | 0.324 | 4.353 | |
KH2PO4 | 0.33 | g | 0.324 | 2.384 | |
MgCl2 x 6 H2O | 0.33 | g | 0.324 | 1.596 | |
CaCl2 x 2 H2O | 0.33 | g | 0.324 | 2.207 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
Sulfur
(powdered) |
10.00 | g | 9.833 | 306.606 | |
Yeast extract | 3.00 | g | 2.95 | - | |
Wolin's vitamin solution (10x) | 1.00 | ml | - | - | |
Neutralized sulfide solution 3% (w/v) | 15.00 | ml | - | - | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients except sulfur, yeast extract, vitamins and sulfide. Adjust pH to 3.5 with H2SO4 and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials containing the appropriate amount of sulfur. Heat vessels containing medium to 90°C for 1 hour on each of 3 succesive days. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The sulfide stock solution (3% w/v) should be neutralized before use (see medium 29). The pH of the complete medium should be at 3.5 - 3.8. |
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] | |
---|---|---|---|---|---|
Na2S x 9 H2O | 3 | g | 30 | 124.905 | |
Distilled water | 100 | ml | - | - | |
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle. |
Last modified: | 08.04.22 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 3.5 - 3.8 |
Equipment needed: | Filter, Gassing station, Hungate tubes, Water bath |
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] |
---|---|---|
Sulfur | 9.833 | 306.606 |
Yeast extract | 2.95 | - |
Na2S x 9 H2O | 0.442 | 1.842 |
NH4Cl | 0.324 | 6.066 |
KCl | 0.324 | 4.353 |
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