| Mineral salt solution* ("Hutner/Cohen-Bazire") | 20 | ml |
| Bacto Peptone (Difco) | 0.25 | g |
| Bacto Yeast Extract (Difco) | 0.25 | g |
| Bacto Agar (Difco) | 15 | g |
| Glucose solution (2.5%, filter-sterilized) | 10 | ml |
| Vitamin solution** | 5 | ml |
| Artificial sea water*** | 250 | ml |
| Distilled water | 715 | ml |
| pH 7.5 | |
|
| | Mix ingredients (except glucose and vitamin solutions), and sterilize 20 min at 121℃. After cooling to 60℃, aseptically add glucose and vitamin solutions to the medium. The medium is only weakly buffered; one needs approximately 10 drops/1 medium of 6 N KOH. |
| | | | | | |
| | *Mineral salt solution |
| | | Nitrilotriacetic acid (NTA) | 10 | g |
| | | MgSO4·7H2O | 29.7 | g |
| | | CaCl2·2H2O | 3.34 | g |
| | | Na2MoO4·2H2O | 12.67 | mg |
| | | FeSO4·7H2O | 99 | mg |
| | | Metal salt solution 44**** | 50 | ml |
| | | Distilled water | 900 | ml |
| | Dissolve NTA first by neutralizing with KOH, then add other salts. Adjust pH to 7.2 with KOH or H2SO4. Adjust volume to 1 L with distilled water. |
| | | | | | |
| | **Vitamin solution |
| | | Biotin | 4 | mg |
| | | Folic acid | 4 | mg |
| | | Pyridoxine-HCl | 20 | mg |
| | | Riboflavin | 10 | mg |
| | | Thiamine-HCl | 10 | mg |
| | | Nicotinamide | 10 | mg |
| | | Ca-pantothenate | 10 | mg |
| | | Vitamin B12 | 0.2 | mg |
| | | p-Aminobenzoic acid | 10 | mg |
| | | Distilled water | 1 | L |
| | | | | | |
| | ***Artificial sea water |
| | | NaCl | 23.477 | g |
| | | Na2SO4 | 3.917 | g |
| | | MgCl2·6H2O | 4.981 | g |
| | | CaCl2 | 1.102 | g |
| | | NaHCO3 | 192 | mg |
| | | KCl | 664 | mg |
| | | KBr | 6 | mg |
| | | H3BO3 | 26 | mg |
| | | SrCl2 | 24 | mg |
| | | NaF | 3 | mg |
| | | Distilled water | 1 | L |
| | | | | | |
| | ****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 | 100 | ml |
|
| | 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℃). |