Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
|Compound||Amount||Unit||Conc. [g/L]||Conc. [mM]|
|Clarified rumen fluid||400.00||ml||-||-|
|CaCl2 x 2 H2O||0.06||g||0.06||0.408|
|MgSO4 x 7 H2O||0.09||g||0.09||0.365|
|L-Cysteine HCl x H2O||0.30||g||0.3||1.708|
|1 Dissolve ingredients (except bicarbonate, cellobiose, yeast extract, cysteine and DTT), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Add the bicarbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add cellobiose, yeast extract, cysteine and DTT from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Cellobiose has to be sterilzed by filtration. Adjust pH of complete medium to 6.7 - 6.8, if necessary.|
|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.|
|Medium type:||Complex medium|
|Final pH:||6.7 - 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|