Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Sea Salt
(SIGMA) |
30.00 | g | 29.851 | - | |
NH4Cl | 1.00 | g | 0.995 | 18.602 | |
KH2PO4 | 0.35 | g | 0.348 | 2.559 | |
MES [2-(N-morpholino) ethane sulfonic acid] | 1.95 | g | 1.94 | 9.938 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Selenite-tungstate solution | 1.00 | ml | - | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
Sulfur , powdered | 10.00 | g | 9.95 | 310.267 | |
Na2CO3 | 0.50 | g | 0.498 | 4.694 | |
Seven vitamins solution | 1.00 | ml | - | - | |
Growth-stimulating factors | 1.00 | ml | - | - | |
Na-dithionite solution (5% w/v) | 1.00 | ml | - | - | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except sulfur, carbonate, vitamins, growth-stimulating factors and Na-dithionite), boil medium for 1 min, then cool to room temperature under 80% H2 and 20% CO2 gas atmosphere. Adjust pH to 6.0 and dispense under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. Autoclave at a temperature of 110°C for 20 min! Add vitamins and growth-stimulating factors from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate and Na-dithionite from sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins and Na-dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.0, if necessary. | |||||
2 After inoculation use 1 bar overpressure of sterile 80% H2 and 20% CO2 gas mixture. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Isobutyric acid | 5 | g | 5 | 56.747 | |
Valeric acid | 5 | g | 5 | 48.957 | |
2-Methylbutyric acid | 5 | g | 5 | 48.957 | |
3-Methylbutyric acid | 5 | g | 5 | 48.956 | |
Caproic acid | 2 | g | 2 | 17.218 | |
Succinic acid | 6 | g | 6 | 50.809 | |
Distilled water | 1000 | ml | - | - | |
1 Dissolve ingredients, adjust pH to 9.0 with NaOH, then autoclave under 100% N2 gas. |
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] |
---|---|---|---|---|
Vitamin B12 | 100 | mg | 0.1 | 0.074 |
p-Aminobenzoic acid | 80 | mg | 0.08 | 0.583 |
D-(+)-biotin | 20 | mg | 0.02 | 0.082 |
Nicotinic acid | 200 | mg | 0.2 | 1.625 |
Calcium pantothenate | 100 | mg | 0.1 | 0.21 |
Pyridoxine hydrochloride | 300 | mg | 0.3 | 1.459 |
Thiamine-HCl x 2 H2O | 200 | mg | 0.2 | 0.536 |
Distilled water | 1000 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaOH
(1 N) |
1.0 | ml | - | - | |
Na2S2O4 | 0.5 | g | 50 | 287.175 | |
Distilled water | 9.0 | ml | - | - | |
1 Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly. |
Last modified: | 14.06.24 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 6.0 |
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] |
---|---|---|
Sea Salt | 29.851 | - |
Sulfur | 9.95 | 310.267 |
MES [2-(N-morpholino) ethane sulfonic acid] | 1.94 | 9.938 |
NH4Cl | 0.995 | 18.602 |
Na2CO3 | 0.498 | 4.694 |
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