f/2 Medium
(Guillard & Ryther 1962, Guillard 1975)
To 950 mL filtered awater add:
太级Quantity | Compound | Stock Solution | Molar Concentration in Final Medium |
1 mL | NaNO3 | 75 g/L dH2O | 8.83 x 10-4 M |
1 mL | NaH2PO4 · H2O | 5 g/L dH2O | 3.63 x 10-5 M |
1 mL * | Na2SiO3 · 9H2O* | 30 g/L dH2O* | 1.07 x 10-4 M* |
1 mL | f/2 trace metal solution | (e recipe below) | - |
0.5 mL | f/2 vitamin solution | (e recipe below) | - |
| | | |
Make final volume up to 1 L with filtered awater and autoclave.
*Note: Autoclaved f/2 medium produces extensive silica precipitate. We delete silicate when it is not required by the alga (e f/2-Si medium below).
f/2 Trace Metal Solution
(Guillard & Ryther 1962, Guillard 1975)
To 950 mL dH2O add:
Quantity | Compound | Stock Solution | Molar Concentration in Final Medium |
3.15 g | FeCl3 · 6H2O | - | 1 x 10-5 M |
4.36 g | Na2EDTA · 2H"在家创业"2O | - | 1 x 10-5 M |
| 游泳圈简笔画 1 mL | CuSO4 · 5H2O 家常素菜100道素菜 | 9.8 g/L dH2O | 4 x 10-8 M |
1 mL | Na2MoO4 · 2H2O | 6.3 g/L dH2O | 3 x 10-8 M |
1 mL | ZnSO4 · 7H2O | 22.0 g/L dH2O | 8 x 10-8 M 米开朗琪罗传 |
1 mL | CoCl2 · 6H2O | 10.0 g/L dH2O | 5 x 10-8 M |
1 mL | MnCl2 · 4H2O | 180.0 g/L dH2O | 9 x 10-7 M |
| | | |
Make final volume up to 1 L with dH2O. Autoclave.
f/2 Vitamin Solution
(Guillard & Ryther 1962, Guillard 1975)
To 950 mL dH2O add:
Quantity | Compound | Stock Solution | Molar Concentration in Final Medium |
1 mL | Vitamin B12 (cyanocobalamin) | 1.0 g/L dH2O | 1 x 10-10 M |
10 mL | Biotin | 0.1 g/L dH2O | 2 x 10-9 M |
200 mg | Thiamine · HCl | - | 3 x 10-7 M |
| | | |
Make final volume up to 1 L with dH2O. Autoclave and store in refrigerator. Note: Vitamin B12 and biotin are obtained in a crystalline form. When preparing the vitamin B12 stock solution, allow for approximately 11% water of crystallization (for each 1 mg of Vitamin B12, add 0.89 mL dH2O). When preparing the biotin stock solution, allow for approximately 4% water of crystallization (for each 1 mg of biotin, add 9.6 mL dH2O).
f/2 Derivatives
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, dispen 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 aptically dispen 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 becau less precipitation forms.
f/2 + Se: Extra silicon and lenium are beneficial to veral diatom strains. Prepare 1 L of f/2 medium but u 2 mL of silicate stock, then add 1.0 mL of lenium 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 awater. Add f/2 medium nutrients and autoclave.
f/2-Si (24 psu): Mix 750 mL distilled H2O and 250 mL filtered awater. Prepare as for f/2 medium but omit Na2适可而止的意思SiO 3 · 9H2O.
f/4: Add 500 mL f/2 medium to 500 mL filtered awater, then autoclave.
f/4-Si: Autoclave 1 L of filtered awater. When cool, aptically add f/2-Si nutrients at half concentration (i.e., 0.5 mL).
f/20-Si: 粉红色的英文Autoclave 1 L of filtered awater. When cool, aptically 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 awater in a Teflon-lined bottle. Wait for the autoclaved awater to cool to room temperature (important). Aptically add 40 μL of sterile f/2 nutrients (20 μL of vitamins).
f/50-Si + CCMP1320 as food: Prepare f/50 and aptically add 50 μL of healthy, moderately den culture of CCMP1320.
f/2m: To 1L f/2 medium add 1 g methylamine · HCl, mix until dissolved and autoclave. This medium is ud to test for contamination by methylaminotrophic bacteria.
f/2p: 萧山萝卜干To 1 L f/2 medium, add 1 g Bacto-peptone, mix until dissolves and autoclave. This medium is ud 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 ud to test for contamination by bacteria and fungi.
f/2 + NPM: Add f/2 nutrients to 900 mL of awater and autoclave. After cooling, aptically add 100 mL of the following organic stock solution. Dispen aptically into test tubes.
