Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 1.50 | g | ||
|
Na2HPO4 | 0.60 | g | ||
|
KCl | 0.10 | g | ||
|
Na-acetate | 0.82 | g | ||
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
NaHCO3 | 2.50 | g | ||
|
Na2-fumarate | 8.00 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients (except bicarbonate, fumarate and vitamins), sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and equilibrate the medium with 80% N2 and 20% CO2 gas mixture to reach a pH of 6.8, then dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add fumarate and vitamins from anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration. Check medium pH and adjust to 6.8, if necessary. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
MnSO4 x H2O | 0.50 | g | ||
|
NaCl | 1.00 | g | ||
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
H3BO3 | 0.01 | g | ||
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
Distilled water | 1000.00 | ml | ||
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. |
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 |