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Medium No.	376
Medium	Ekho Lake Strains Medium
Composition
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℃).
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