Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 30.00 | g | ||
|
KCl | 0.20 | g | ||
|
K2HPO4 | 0.20 | g | ||
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
NH4Cl | 1.00 | g | ||
|
Yeast extract | 0.50 | g | ||
|
Peptone | 5.00 | g | ||
|
Trace element solution (mg per 200 ml) | 1.00 | ml | ||
|
Resazurin | 1.00 | mg | ||
|
Distilled water | 1000.00 | ml | ||
1 Prepare the medium omitting Na2CO3, NaHCO3, Na2S x 9 H2O, and Vitamin solution. | |||||
2 Boil the medium and cool under nitrogen, add Na2CO3 and NaHCO3. | |||||
|
NaHCO3 | 24.00 | g | ||
|
Na2CO3 | 2.76 | g | ||
3 Adjust pH to 9.5-10.0 with 10M KOH. | |||||
4 Dispense into tubes or bottle fitted with rubber stoppers. | |||||
5 Autoclave at 121 °C for 15 min. | |||||
6 Add Na2S x 9 H2O (neutralized) and Vitamin solution from sterile stock solutions | |||||
|
Na2S x 9 H2O
(from neutralized stock solution) |
0.40 | g | ||
|
Wolin's vitamin solution | 2.00 | ml |
Compound | Amount | Unit | |
---|---|---|---|
|
MnCl2 x 4 H2O | 720 | mg |
|
Fe(NH4)2(SO4)2 x 12 H2O | 400 | mg |
|
FeSO4 x 7 H2O | 200 | mg |
|
CoCl2 x 6 H2O | 200 | mg |
|
ZnSO4 x 7 H2O | 200 | mg |
|
Na2MoO4 x 2 H2O | 20 | mg |
|
NiCl2 | 100 | mg |
|
CuSO4 x 5 H2O | 20 | mg |
|
AlK(SO4)2 x 12 H2O | 20 | mg |
|
H3BO3 | 20 | mg |
|
HCl
(concentrated) |
5 | ml |
|
Distilled water | 200 | 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 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 |