Reference: Cowgill, U.M. and Milazzo, D.P. 1989. Aquatic Toxicology and Hazard Assessment. 12th volume. ASTM STP 1027. pp. 379-391.
| Stock | Substance | Concentration | Amount added to medium |
|---|---|---|---|
| 1. |
MgSO4•7H2O |
24.6 g/100 mL |
2 mL/L |
| 2. | Ca(NO3)2•4H2O | 23.6 g/100 mL |
5 mL/L |
| 3. | KH2PO4 | 13.6 g/100 mL |
5 mL/L |
| 4. |
KNO3 |
10.1 g/100 mL | 10 mL/L |
| 5. | Fe-Na2EDTA•2H2O* | See below |
20 mL/L |
| 6. |
H3BO3 |
2.86 g/L |
1 mL/L |
| ZnSO4•7H2O |
0.22 g/L |
||
| Na2MoO4•2H2O |
0.12 g/L |
||
| CuSO4•5H2O | 0.08 g/L | ||
| MnCl2•4H2O | 3.62 g/L | ||
| 7. | Sucrose | – |
10.00 g/L |
| 8. | Yeast extract | – |
0.10 g/L |
| 9. |
Bacto-tryptone or Bacto-peptone |
– | 0.60 g/L |
For one litre of medium start with about 900 mL of purified water and add each stock solution in the order above. The medium is then stirred until all the contents are dissolved. Adjust the pH to 4.60 with NaOH or HCl and bring volume up to 1 L with purified water and autoclave. Stock solutions should be stored in the dark (i.e., dark amber or aluminum-covered bottles) due to potential photosensitivity. Stock solutions may be stored in the refrigerator for up to 6 months, provided that they are isolated from solvents or other potential contaminants. Monitor the Fe-Na2EDTA solution for precipitate.
* Fe-Na2EDTA solution:
To 1000 mL of purified water add 1.5 g of Na2EDTA•2H2O and dissolve completely. Then add 0.48 g of FeCl3•6H2O and dissolve. This solution may be added directly together with the above stock solutions and autoclaved together with the medium. Alternatively, if medium forms precipitate then autoclave this solution separately and add aseptically after autoclaving both solutions.