F/2 Culture Media
F/2 Medium Stock Solutions:
|
Quantity per L of Media |
Compound |
Stock Concentration |
|
| 1.0 ml | NaNO3 | 75.0 g/L dH2O | Autoclave separately |
| 1.0 ml | NaH2PO4.H2O | 5.0 g/L dH2O | Autoclave separately |
| 1.0 ml | Na2SiO3.9H2O | 30.0 g/L dH2O | Autoclave separately |
| 1.0 ml | f/2 Trace Metal Solution | (see recipe below) | Filter sterilize |
| 0.5 ml | f/2 Vitamin Solution | (see recipe below) | Filter sterilize |
Stock solutions (except vitamins) are considered 1000x. Seawater should be autoclaved and cooled prior to mixing the media. We tend to sterile-filter artificial seawater rather than autoclaving to avoid reductions in dissolved carbon dioxide. A
Add all the f/2 components aseptically to your media once it has cooled to room temperature. For other algal groups (e.g. dinoflagellates, coccolithophores) use f/2-Si medium (prepare as for f/2 medium but omit Na2SiO3.9H2O). Sterile stock solutions can be stored at room temperature. Once opened, stock solutions should be stored at 4 °C.
Trace Metal Solution
To 950 mL distilled H2O add:
|
Quantity |
Compound |
Stock Solution |
| 1.3 g | FeCl3.6H2O | |
| 8.7 g | Na2EDTA.2H2O | |
| 1.0 ml | CuSO4.5H2O | 980 mg / 100 ml dH20 |
| 1.0 ml | Na2MoO4.2H2O | 630 mg / 100 ml dH20 |
| 1.0 ml | ZnSO4.7H2O | 2.2 g / 100 ml dH20 |
| 1.0 ml | CoCl2.6H2O | 1.0 g / 100 ml dH20 |
| 1.0 ml | MnCl2.4H2O | 18.0 g / 100 ml dH20 |
Initially, the solution will be cloudy. Add 1N NaOH to adjust pH to about 4.5 to clear the solution. Bring final volume to one liter.
Vitamin Solution
To 950 mL dH2O add:
|
Quantity |
Compound |
1o Stock Solution |
Tip |
| 1.0 ml | Vitamin B12 | 10 mg / 10 ml dH2O | |
| 1.0 ml | Biotin | 10 mg / 10 ml dH2O | Dissolve in a little 0.1N NaOH first |
| 200.0 mg | Thiamine HCl |
Make final volume up to 1.0 L with dH2O. Store primary stock solutions in freezer. Store completed vitamin solution in the refrigerator or freeze small aliquots.
References
Guillard R.R.L. and Ryther J.H. (1962) Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea Cleve. Can. J. Microbiol. 8: 229-239.
Guillard R.R.L. (1975) Culture of phytoplankton for feeding marine invertebrates. in “Culture of Marine Invertebrate Animals.” (eds: Smith W.L. and Chanley M.H.) Plenum Press, New York, USA. pp 26-60.