Black Sea Medium: For brackish water organisms (16 psu, half-strength nutrients). Combine 500 mL f/2 medium and 500 mL dH2O. Autoclave.
f/2 agar: Prepare 1 liter of f/2 medium and dissolve 9g Bacto-agar (heat and mix). For test tubes, dispense dissolved agar medium into tubes, autoclave, and then cool with tubes slanted at an angle. For Petri plates, autoclave in a flask, cool almost to the gelling point, and then aseptically dispense into sterile Petri plates. Note: Agar can be added to other media (e.g., f/50 agar), and agar concentration can be varied to produce softer or firmer substrates.
f/2-Si: Prepare as for f/2 medium but omit Na2SiO 3 · 9H2O. This is preferred over f/2 medium for organisms with no silica requirement because less precipitation forms.
f/2 + Se: Extra silicon and selenium are beneficial to several diatom strains. Prepare 1 L of f/2 medium but use 2 mL of silicate stock, then add 1.0 mL of selenium stock solution (1.29 mg H2SeO 3 /L distilled H2O). Autoclave.
f/2 (11 psu): For brackish water organisms. Mix 650 mL distilled H2O and 350 mL filtered seawater. Add f/2 medium nutrients and autoclave.
f/2-Si (24 psu): Mix 750 mL distilled H2O and 250 mL filtered seawater. Prepare as for f/2 medium but omit Na2SiO 3 · 9H2O.
f/4: Add 500 mL f/2 medium to 500 mL filtered seawater, then autoclave.
f/4-Si: Autoclave 1 L of filtered seawater. When cool, aseptically add f/2-Si nutrients at half concentration (i.e., 0.5 mL).
f/20-Si: Autoclave 1 L of filtered seawater. When cool, aseptically add f/2-Si nutrients at one tenth concentration (i.e., 100 μL).
f/50-Si: This is more than a 1/25 dilution of f/2-Si medium. We autoclave 1 L of seawater in a Teflon-lined bottle. Wait for the autoclaved seawater to cool to room temperature (important). Aseptically add 40 μL of sterile f/2 nutrients (20 μL of vitamins).
f/50-Si + CCMP1320 as food: Prepare f/50 and aseptically add 50 μL of healthy, moderately dense culture of CCMP1320.
f/2m: To 1L f/2 medium add 1 g methylamine · HCl, mix until dissolved and autoclave. This medium is used to test for contamination by methylaminotrophic bacteria.
f/2p: To 1 L f/2 medium, add 1 g Bacto-peptone, mix until dissolves and autoclave. This medium is used to test for contamination by non- methylaminotrophic bacteria and fungi.
f/2pm: To 1L f/2 medium add 1 g Bacto-peptone and 1 g methylamine · HCl, mix until dissolved and autoclave. This general medium is used to test for contamination by bacteria and fungi.
f/2 + NPM: Add f/2 nutrients to 900 mL of seawater and autoclave. After cooling, aseptically add 100 mL of the following organic stock solution. Dispense aseptically into test tubes.
Organics Stock Solution
(modified from Guillard 1960)
To 900 mL dH2O add:
|1 g||sodium acetate|
|3 g||(di-) sodium succinate · 6H2O|
|100 mg||yeast extract|
Bring up to 1 L with dH2O. Dispense in small aliquots and autoclave.
Guillard, R.R.L. 1960. A mutant of Chlamydomonas moewusii lacking contractile vacuoles. J. Protozool. 7: 262-268.
Guillard, R.R.L. 1975. Culture of phytoplankton for feeding marine invertebrates. pp 26-60. In Smith, W.L. and Chanle,y M.H. (eds.) Culture of Marine Invertebrate Animals. Plenum Press, New York, USA.
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.