Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 0.50 | g | ||
|
MgSO4 x 7 H2O | 1.90 | g | ||
|
MgCl2 x 6 H2O | 1.40 | g | ||
|
K2HPO4 | 1.60 | g | ||
|
Na2HPO4 x 2 H2O | 1.00 | g | ||
|
NaCl | 26.00 | g | ||
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
Yeast extract
(OXOID) |
2.00 | g | ||
|
Trypticase peptone
(BD BBL) |
2.00 | g | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Starch
(soluble) |
5.00 | g | ||
|
CaCl2 x 2 H2O | 0.56 | 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 starch, calcium chloride, vitamins and reducing agents) and adjust pH to 6.5 - 6.8. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (to 30% of volume) and autoclave. Add starch, calcium chloride, vitamins (sterilized by filtration), sulfide and cysteine from sterile anoxic stock solutions prepared under 100 N2 gas. Adjust pH of complete medium 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 |