Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
KH2PO4 | 0.10 | g | ||
|
(NH4)2SO4 | 0.10 | g | ||
|
MgSO4 x 7 H2O | 0.10 | g | ||
|
CaCl2 x 2 H2O | 0.02 | g | ||
|
Hutner's salts | 1.00 | ml | ||
|
Vitamin solution | 1.00 | ml | ||
|
Yeast extract | 0.10 | g | ||
|
Glucose | 0.10 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Prepare the liquid medium without Staley´s vitamin solution and glucose. Adjust to pH 4.8 - 5.5. Dispense the medium into serum bottles. The gas phase is air:CO2 (95:5) and the liquid:gas phase should be 1:9. Sterilize by autoclaving. After autoclaving add the Staley´s vitamin solution (filter sterilised) and glucose (autoclaved) from sterile stock solutions. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Nitrilotriacetic acid
(NTA) |
10.000 | g | ||
|
MgSO4 x 7 H2O | 29.700 | g | ||
|
CaCl2 x 2 H2O | 3.335 | g | ||
|
(NH4)6Mo7O24 x 4 H2O | 9.250 | mg | ||
|
FeSO4 x 7 H2O | 99.000 | mg | ||
|
"Metals 44" | 50.000 | ml | ||
|
Distilled water | 950.000 | ml | ||
1 Dissolve the nitrilotriacetic acid. | |||||
2 Adjust pH to 7.0 with KOH (~7.3 g). | |||||
3 Dissolve other salts separately and combine. | |||||
4 Adjust pH to 6.8 with NaOH or H2SO4. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Na-EDTA | 250.0 | mg | ||
|
ZnSO4 x 7 H2O | 1095.0 | mg | ||
|
FeSO4 x 7 H2O | 500.0 | mg | ||
|
MnSO4 x H2O | 154.0 | mg | ||
|
CuSO4 x 5 H2O | 39.2 | mg | ||
|
Co(NO3)2 x 6 H2O | 24.8 | mg | ||
|
Na2B4O7 x 10 H2O | 17.7 | mg | ||
|
Distilled water | 1000.0 | ml | ||
1 Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Biotin | 4.0 | mg | ||
|
Folic acid | 4.0 | mg | ||
|
Pyridoxine hydrochloride | 20.0 | mg | ||
|
Riboflavine | 10.0 | mg | ||
|
Thiamine-HCl x 2 H2O | 10.0 | mg | ||
|
Nicotinamide | 10.0 | mg | ||
|
D-Calcium pantothenate | 10.0 | mg | ||
|
Vitamin B12 | 0.2 | mg | ||
|
p-Aminobenzoic acid | 10.0 | mg | ||
|
Distilled water | 1000.0 | ml | ||
1 Filter-sterilized. |