Artificial Seawater Medium



Artificial Seawater Medium Recipe

Modification of Brand's ASP2 medium for 2268 Agmenellum quadruplicatum; suitable for other axenic marine cultures (salinity = 18 ppt).

Directions

For 1 L Total

    1. To approximately 850 mL of dH2O, add each of the components in the order specified (except vitamins) while stirring continuously.
    2. Adjust the pH to 8.1
    3. Bring the total volume to 1 L with dH2O.
      • *For 1.5% agar medium: add 15 g of agar to the flask; do not mix.
    4. Cover and autoclave medium.
    5. Allow to cool and add vitamin B12.
      • *For agar medium: add vitamin, mix, and dispense before agar solidifies.
    6. Store at refrigerator temperature.
# Component Amount Stock Solution Concentration Final Concentration
1 NaCl (Fisher S271-500) 18 g/L   0.31 M
2 MgSO4·7H2O (Sigma 230391) 10 mL/L 26 g/100 mL dH20 10.5 mM
3 KCl (Fisher P 217) 10 mL/L 6 g/100 mL dH20 8 mM
4 NaNO3 (Fisher BP360-500) 10 mL/L 10 g/100 mL dH20 11.8 mM
5 CaCl2·2H2O (Sigma C-3881) 10 mL/L 3 g/100 mL dH20 2 mM
6 KH2PO4 (Sigma P 0662) 10 mL/L 0.5 g/100 mL dH20 0.37 mM
7 Tricine (adjust to pH 8) (Sigma T-5816-256) 20 mL/L 22.4 g/100 mL dH20 25 mM
8 P-II Metal Solution 10 mL/L    
9 Chelated Iron Solution 1 mL/L    
10 NH4Cl (Fisher A 649-500) 1 mL/L 2.7 g/100 mL dH20 0.5 mM
11 Vitamin B12 1 mL/L