Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
K2HPO4 | 0.23 | g | ||
|
KH2PO4 | 0.23 | g | ||
|
NaCl | 0.45 | g | ||
|
(NH4)2SO4 | 0.45 | g | ||
|
CaCl2 x 2 H2O | 0.06 | g | ||
|
MgSO4 x 7 H2O | 0.09 | g | ||
|
Yeast extract | 5.00 | g | ||
|
Indigocarmine | 5.00 | mg | ||
|
NaHCO3 | 6.40 | g | ||
|
D-Glucose | 5.00 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
L-Cysteine HCl x H2O | 0.30 | g | ||
|
Na2S x 9 H2O | 0.30 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients (except bicarbonate, vitamins, glucose, cysteine and sulfide), then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the bicarbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute medium under CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add vitamins, glucose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins are sterilized by filtration. Adjust pH of complete medium to 6.7 - 6.8, if necessary. |
Compound | Amount | Unit | |
---|---|---|---|
|
Biotin | 20 | mg |
|
Folic acid | 20 | mg |
|
Pyridoxine hydrochloride | 100 | mg |
|
Thiamine HCl | 50 | mg |
|
Riboflavin | 50 | mg |
|
Nicotinic acid | 50 | mg |
|
Calcium D-(+)-pantothenate | 50 | mg |
|
Vitamin B12 | 1 | mg |
|
p-Aminobenzoic acid | 50 | mg |
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
Distilled water | 1000 | ml |