Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
KH2PO4 | 0.75 | g | ||
|
K2HPO4 | 1.50 | g | ||
|
NaCl | 0.90 | g | ||
|
MgCl2 x 6 H2O | 0.20 | g | ||
|
NH4Cl | 0.90 | g | ||
|
Modified Wolin's mineral solution | 9.00 | ml | ||
|
Tryptone
(BD Bacto) |
2.00 | g | ||
|
Yeast extract
(OXOID) |
2.00 | g | ||
|
FeSO4 x 7 H2O solution (0.1% w/v) | 0.50 | ml | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Sodium citrate | 2.00 | g | ||
|
Wolin's vitamin solution | 5.00 | ml | ||
|
Na2S x 9 H2O | 0.30 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients (except ferrous sulfate, citrate, vitamins and sulfide), adjust pH to 6.5 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add ferrous sulfate and dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add citrate, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and adjust pH of complete medium to 7.0, 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 | 2.0 | mg |
|
Folic acid | 2.0 | mg |
|
Pyridoxine hydrochloride | 10.0 | mg |
|
Thiamine HCl | 5.0 | mg |
|
Riboflavin | 5.0 | mg |
|
Nicotinic acid | 5.0 | mg |
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
Vitamin B12 | 0.1 | mg |
|
p-Aminobenzoic acid | 5.0 | mg |
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
Distilled water | 1000.0 | ml |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
FeSO4 x 7 H2O | 1 | g | ||
|
H2SO4
(0.1 N) |
1000 | ml | ||
1 The ferrous sulfate solution is not stable and should be freshly prepared. |