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 | 30 | ml | - | - | |
Solution C | 1 | ml | - | - | |
Solution D | 25 | ml | - | - | |
Solution E | 10 | ml | - | - | |
Solution F | 10 | ml | - | - | |
Solution G | 10 | ml | - | - | |
1 Dissolve ingredients of solution A, adjust pH to 7.0, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is filter-sterilized and then equilibrated with 80% N2 and 20% CO2 gas mixture for at least 15 min. Solution C is prepared under 100% N2 gas and sterilized by filtration. Solutions D, E, F and G are autoclaved under 100% N2 gas. To complete the medium, appropriate amounts of the solutions B to G are added to solution A in the sequence indicated. Adjust pH of complete medium to 7.2, if necessary. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
KH2PO4 | 0.50 | g | 0.543 | 3.989 |
MgCl2 x 6 H2O | 0.33 | g | 0.358 | 1.762 |
NaCl | 0.40 | g | 0.434 | 7.432 |
NH4Cl | 0.40 | g | 0.434 | 8.119 |
CaCl2 x 2 H2O | 0.05 | g | 0.054 | 0.369 |
Trace element solution SL-10 | 1.00 | ml | - | - |
Clarified rumen fluid | 50.00 | ml | - | - |
Trypticase peptone
(BD BBL) |
1.00 | g | 1.086 | - |
PIPES
(SIGMA) |
15.00 | g | 16.287 | 53.863 |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.4e-4 | 0.002 |
Distilled water | 870.00 | 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-laurate | 2.78 | g | 111.2 | 500.225 |
Distilled water | 25.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
CaCl2 x 2 H2O | 1.84 | g | 184 | 1251.57 |
Distilled water | 10.00 | ml | - | - |
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: | 10.10.22 |
Source: | DSMZ |
Taxonomic range: | Bacteria |
Medium type: | Complex medium |
Final pH: | 7.2 |
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] |
---|---|---|
PIPES | 14.896 | 49.263 |
Na-laurate | 2.761 | 12.419 |
CaCl2 x 2 H2O | 1.827 | 12.429 |
Na2CO3 | 1.49 | 14.054 |
Trypticase peptone | 0.993 | - |
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