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Artificial Seawater Medium

Directions

Modification of Brand's ASP2 medium for 2268 Agmenellum quadruplicatum; suitable for other axenic marine cultures (salinity = 30 ppt).

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 M
3	KCl (Fisher P 217)	10 mL/L	6 g/100 mL dH20	8 M
4	NaNO₃ (Fisher BP360-500)	10 mL/L	10 g/100 mL dH20	11.8 M
5	CaCl₂·2H₂O (Sigma C-3881)	10 mL/L	3 g/100 mL dH20	2 M
6	KH₂PO₄ (Sigma P 0662)	10 mL/L	0.5 g/100 mL dH20	0.29 M
7	Tricine (adjust to pH 8) (Sigma T-5816-256)	20 mL/L	22.4 g/100 mL dH20	25 M
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 M
11	Vitamin B₁₂	1 mL/L

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