Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Solution A | 921 | ml | - | - | |
Solution B | 50 | ml | - | - | |
Solution C | 1 | ml | - | - | |
Solution D | 25 | ml | - | - | |
Solution E | 10 | ml | - | - | |
Solution F | 1 | ml | - | - | |
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.4 and dispense 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 atmosphere and sterilized by filtration. Solutions D, E and F are autoclaved separately under 100% N2 gas. To complete the medium add appropriate amounts of solutions B - F to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 6.4 - 6.8, if necessary. | |||||
2 Note: Rumen fluid may be replaced by supplementing the medium with 200 µg/ml of crude egg yolk phosphatidylcholine (SIGMA, type IX-E). |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Mineral solution | 50.0 | ml | - | - |
Trace element solution SL-10 | 1.0 | ml | - | - |
Clarified rumen fluid | 300.0 | ml | - | - |
Trypticase peptone
(BD Bacto/BD BBL) |
5.0 | g | 5.429 | - |
Na-formate | 0.6 | g | 0.651 | 9.579 |
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.4e-4 | 0.002 |
Distilled water | 570.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
NaHCO3 | 2.5 | g | 50 | 595.188 |
Distilled water | 50.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Lactose | 5 | g | 200 | 584.283 |
Distilled water | 25 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
L-Cysteine HCl x H2O | 0.24 | g | 24 | 136.646 |
Distilled water | 10.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2S x 9 H2O | 78 | mg | 78 | 324.754 |
Distilled water | 1 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
KH2PO4 | 10.0 | g | 10 | 73.484 |
MgCl2 x 6 H2O | 6.6 | g | 6.6 | 32.464 |
NaCl | 8.0 | g | 8 | 136.893 |
NH4Cl | 8.0 | g | 8 | 149.561 |
CaCl2 x 2 H2O | 1.0 | g | 1 | 6.802 |
Distilled water | 1000.0 | 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: | 10.10.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 6.4 - 6.8 |
Equipment needed: | Autoclave, Centrifuge, 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 .
All strains for this medium: | 1 |
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Lactose | 4.96 | 14.491 |
Trypticase peptone | 4.96 | - |
NaHCO3 | 2.48 | 29.523 |
Na-formate | 0.595 | 8.753 |
KH2PO4 | 0.496 | 3.645 |
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