Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
(NH4)2SO4 | 1.30 | g | ||
|
MgCl2 x 6 H2O | 2.60 | g | ||
|
KH2PO4 | 1.43 | g | ||
|
K2HPO4 | 5.50 | g | ||
|
CaCl2 x 2 H2O | 0.13 | g | ||
|
Na2-ß-glycerophosphate x 5 H2O
(MERCK 35675) |
6.00 | g | ||
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
1.10 | ml | ||
|
L-Glutathione
(reduced) |
0.25 | g | ||
|
Yeast extract | 4.50 | g | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Cellobiose | 5.00 | g | ||
|
Cellulose
(Avicel or MN 301) optional |
10.00 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except cellobiose, sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then adjust pH to 7.0 - 7.2, distribute under same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Cellobiose is added to the sterile medium from an anoxic 10% (w/v) stock solution prepared under 100% N2 gas and sterilized by filtration. Some strains can be adapted to cellulose as substrate using 10.00 g/l cellulose (Avicel or MN 301, MACHEREY-NAGEL). | |||||
2 Note: A white precipitate forms after mixing the ingredients of this medium, but this has no negative effect on growth. |