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Main sol. 330

Internal Solution-ID: S606

Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Mineral solution 38.00 ml - -
K2HPO4 0.30 g 0.3 1.722
Trypticase peptone
(BD BBL)
2.00 g 2 -
Yeast extract
(OXOID)
0.50 g 0.5 -
Volatile fatty acid mixture 3.10 ml - -
Haemin solution
(0.05% w/v)
2.00 ml - -
Glycerol 0.50 g 0.5 5.429
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Na2CO3 4.00 g 4 37.74
D-Glucose 0.50 g 0.5 2.775
Maltose 0.50 g 0.5 1.461
Cellobiose 0.50 g 0.5 1.461
Starch , soluble 0.50 g 0.5 -
L-Cysteine HCl x H2O 0.25 g 0.25 1.423
Na2S x 9 H2O 0.25 g 0.25 1.041
Distilled water 960.00 ml - -
1 Dissolve ingredients (except carbonate, glucose, maltose, cellobiose, soluble starch, cysteine and sulfide), then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the carbonate 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 glucose, maltose, cellobiose, soluble starch, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 6.7 - 6.8, if necessary.
330 RUMEN BACTERIA MEDIUM