Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Solution A | 930 | ml | - | - | |
| Solution B | 30 | ml | - | - | |
| Solution C | 1 | ml | - | - | |
| Solution D | 20 | ml | - | - | |
| Solution E | 10 | ml | - | - | |
| Solution F | 10 | ml | - | - | |
| 1 Add and dissolve ingredients of solution A, adjust pH to 7.2, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas and sterilized by filtration. Solutions D, E and F are autoclaved under 100% N2 gas. To complete the medium, appropriate amounts of solutions B to F are added to the sterile solution A. Adjust pH of complete medium to 7.2, if necessary. | |||||
| 2 Note: Use 25 - 50% inoculum. Growth can be followed by the clearing of stearate. Maintain cultures at 25°C. | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| KH2PO4 | 0.50 | g | 0.538 | 3.951 |
| MgCl2 x 6 H2O | 0.33 | g | 0.355 | 1.745 |
| NaCl | 0.40 | g | 0.43 | 7.36 |
| NH4Cl | 0.40 | g | 0.43 | 8.041 |
| CaCl2 x 2 H2O | 0.05 | g | 0.054 | 0.366 |
| Trace element solution SL-10 | 1.00 | ml | - | - |
| Clarified rumen fluid | 50.00 | ml | - | - |
|
Trypticase peptone
(BD BBL) |
1.00 | g | 1.075 | - |
| Na2SO4 | 2.80 | g | 3.011 | 21.197 |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 |
| Distilled water | 880.00 | ml | - | - |
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| Na2CO3 | 1.5 | g | 50 | 471.752 |
| Distilled water | 30.0 | ml | - | - |
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Na-stearate | 0.61 | g | 30.5 | 99.524 | |
| Distilled water | 20.00 | ml | - | - | |
| 1 Dissolve stearate by heating in water bath. | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| L-Cysteine HCl x H2O | 0.3 | g | 30 | 170.808 |
| Distilled water | 10.0 | ml | - | - |
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| Na2S x 9 H2O | 0.3 | g | 30 | 124.905 |
| Distilled water | 10.0 | ml | - | - |
| 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. | |||||
| 1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C. |
| 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 | - | - |
| Last modified: | 28.02.22 |
| Source: | DSMZ |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | 7.2 |
| 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 .