Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
KCl | 0.33 | g | ||
|
MgCl2 x 6 H2O | 6.00 | g | ||
|
MgSO4 x 7 H2O | 1.00 | g | ||
|
NH4Cl | 0.25 | g | ||
|
CaCl2 x 2 H2O | 0.14 | g | ||
|
K2HPO4 | 0.14 | g | ||
|
NaCl | 18.00 | g | ||
|
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v) |
2.00 | ml | ||
|
Yeast extract
(Difco) |
1.00 | g | ||
|
Resazurin | 1.00 | mg | ||
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
NaHCO3 | 2.00 | g | ||
|
Glucose | 10.00 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except cysteine, bicarbonate, glucose and vitamins, then sparge medium with 100% N2 gas to make it anoxic. Add cysteine and sodium bicarbonate, then adjust pH to 7.5. Dispense medium in anoxic Hungate-type tubes under 100% N2 gas and autoclave. A precipitate forms which will redissolve slowly over about 3 days. Before use inject separately sterile glucose solution (20% w/v) and vitamins prepared under 100% N2 gas atmosphere. Vitamins should be sterilized by filtration. Check medium pH and adjust pH to 7.6, 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 |