| Peptone | 0.15 | g |
| Yeast extract | 0.15 | g |
| Glucose | 1 | g |
| (NH4)2SO4 | 0.25 | g |
| Vitamin solution* | 10 | ml |
| Hutner's basal salts** | 20 | ml |
| Distilled water | 1 | L |
| pH 7.2 | |
|
| | Autoclave at 121°C for 15 min. Glucose and vitamin solutions should be filter-sterilized separately. |
| | | | | | |
| | *Vitamin solution |
| | | Vitamin B12 | 0.1 | mg |
| | | Biotin | 2 | mg |
| | | Thiamine-HCl | 5 | mg |
| | | Ca-pantothenate | 5 | mg |
| | | Folic acid | 2 | mg |
| | | Riboflavin | 5 | mg |
| | | Nicotinamide | 5 | mg |
| | | Distilled water | 1 | L |
| | | | | | |
| | **Hutner's basal salts |
| | | Nitrilotriacetic acid (NTA) | 10 | g |
| | | MgSO4·7H2O | 29.7 | g |
| | | CaCl2·2H2O | 3.34 | g |
| | | (NH4)6MoO7O24·4H2O | 9.25 | mg |
| | | FeSO4·7H2O | 99 | mg |
| | | Metal salt solution 44*** | 50 | ml |
| | | Distilled water | 950 | ml |
| | Dissolve and neutralized nitrilotriacetic acid in 500 ml distilled water with the addition of KOH. Add the remaining chemicals in order listed. Adjust pH to 6.8, and bring volume to 1 L with distilled water. |
| | | | | | |
| | ***Metal salt solution 44 |
| | | Na-EDTA | 250 | mg |
| | | ZnSO4·7H2O | 1095 | mg |
| | | FeSO4·7H2O | 500 | mg |
| | | MnSO4·H2O | 154 | mg |
| | | CuSO4·5H2O | 39.2 | mg |
| | | Co(NO3)2·6H2O | 24.8 | mg |
| | | Na2B4O7·10H2O | 17.7 | mg |
| | | Distilled water | 1 | L |
|
| | Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions. Store in the dark and cold (5℃). |