Main sol. 159
Internal Solution-ID: S275
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Clarified rumen fluid | 300.00 | ml | - | - | |
CaCl2 x 2 H2O | 0.12 | g | 0.12 | 0.816 | |
MgSO4 x 7 H2O | 0.18 | g | 0.18 | 0.73 | |
KH2PO4 | 0.45 | g | 0.45 | 3.307 | |
K2HPO4 | 0.45 | g | 0.45 | 2.584 | |
NaCl | 0.90 | g | 0.9 | 15.4 | |
(NH4)2SO4 | 0.90 | g | 0.9 | 6.811 | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
L-Cysteine HCl x H2O | 1.00 | g | 1 | 5.694 | |
Na2CO3 | 1.50 | g | 1.5 | 14.153 | |
D-Glucose | 2.00 | g | 2 | 11.101 | |
Distilled water | 700.00 | ml | - | - | |
1 Dissolve ingredients (except cysteine, carbonate and glucose) and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH with KOH to 6.5. Distribute medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Therafter, add glucose from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Final pH of the medium should be 6.7 - 7.0. |
159 | TREPONEMA MEDIUM |