Digestion Medium Preparation and Recipe

Table 1. Required components

Component	Amount	Concentration
Sodium chloride (mw: 58.44 g/mol)	23.4 g	0.4 M
Magnesium Chloride Hexahydrate (mw: 203.30 g/mol)	26.4 g	0.13 M
Magnesium Sulfate Heptahydrate (mw: 246.47 g/mol)	5.4 g	0.022 M
Potassium Chloride (mw: 74.55 g/mol)	11.9 g	0.16 M
Calcium Chloride (mw: 110.98 g/mol)	0.3 g	0.002 M
2-(N-morpholino)ethanesulfonic acid (mw: 195.24 g/mol)	1.95 g	0.01 M

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 23.4 g of Sodium chloride to the solution.
3. Add 26.4 g of Magnesium Chloride Hexahydrate to the solution.
4. Add 5.4 g of Magnesium Sulfate Heptahydrate to the solution.
5. Add 11.9 g of Potassium Chloride to the solution.
6. Add 0.3 g of Calcium Chloride to the solution.
7. Add 1.95 g of 2-(N-morpholino)ethanesulfonic acid to the solution.
8. Add distilled water until the volume is 1 L.
9. Adjust the pH to 6.5. Filter through a 0.2-µm filter and store at 4°C. Just before use, add 1% (w/v) cellulose (Sigma) and 3 U/mL alginate lyase (Boyen et al. 1990).

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