Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Mineral solution 1 | 37.5 | ml | ||
|
Mineral solution 2 | 37.5 | ml | ||
|
(NH4)2Ni(SO4)2 x 6 H2O | 3.0 | mg | ||
|
FeSO4 x 7 H2O | 2.0 | mg | ||
|
Yeast extract
(BD-Difco) |
2.0 | g | ||
|
Trypticase peptone
(BD-BBL) |
2.0 | g | ||
|
Trace vitamins | 10.0 | ml | ||
|
Trace minerals | 10.0 | ml | ||
|
Na2SO4 | 3.4 | g | ||
|
NaHCO3 | 2.0 | g | ||
|
Resazurin | 1.0 | mg | ||
|
Na2S x 9 H2O | 0.5 | g | ||
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
Distilled water | 920.0 | ml | ||
1 Mix ingredients, except NaHCO3, L-cysteine x HCl x H2O and Na2S x 9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20), and add NaHCO3 to the medium. Dispense the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight and adjust pH to 6.5, if necessary. Separately autoclave L-cysteine x HCl and Na2S x 9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the L-cysteine x HCl and Na2S x 9H2O solutions. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20). |
Compound | Amount | Unit | |
---|---|---|---|
|
KH2PO4 | 6.0 | g |
|
(NH4)2SO4 | 6.0 | g |
|
NaCl | 12.0 | g |
|
MgSO4 x 7 H2O | 2.4 | g |
|
CaCl2 x 2 H2O | 1.6 | g |
|
Distilled water | 1000.0 | ml |
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 pantothenate | 5.0 | mg |
|
Vitamin B12 | 0.1 | mg |
|
p-Aminobenzoic acid | 5.0 | mg |
|
Lipoic acid | 5.0 | mg |
|
Distilled water | 1000.0 | ml |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
MnSO4 x n H2O | 0.50 | g | ||
|
NaCl | 1.00 | g | ||
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
CoSO4 x 7 H2O | 0.10 | g | ||
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
AlK(SO4)2 | 0.01 | g | ||
|
H3BO3 | 0.01 | g | ||
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. |