F2 Medium - IBEMicrobeHub/culturing GitHub Wiki
Materials:
- 993,5 mL of Artificial seawater.
- dH2O.
- Filters 0,22 µm.
Table 1
| F/2 medium | Conc.[g/L] | g in 100 mL / 50 mL of stock | Quantity for 1L of medium |
|---|---|---|---|
| Nitrate (NaNO3) | 75,00 g/L | 7,500 g / 3,75 g | 1,0 mL |
| Phosphate (NaH2PO4·H2O) | 5,00 g/L | 0,500 g / 0,25 g | 1,0 mL |
| Silicate (Na2SiO3·9H2O) | 30,00 g/L | 3,000 g / 1,5 g | 1,0 mL |
| FeCl3 (FeCl3·6H2O) | 3,15 g/L | 0,315 g / 0,1575 g | 1,0 mL |
| NaEDTA (NaEDTA·2H2O) | 4,36 g/L | 0,436 g / 0,218 g | 1,0 mL |
| Trace metal | See recipe below (1) | --- | 1,0 mL |
| Vitamins | See recipe below (2) | --- | 0,5 mL |
| Artificial seawater | 36 g/L | --- | 993,5 mL |
*(1) F/2 Trace Metal solution:
Table 2
| Trace metal solution | Conc.[g/L] | g in 100 mL / 50 mL of stock |
|---|---|---|
| MnCl·4H2O | 0,1800 g/L | 0,01800 g / 0,009000 g |
| ZnSO4·7H2O | 0,0220 g/L | 0,00220 g / 0,001100 g |
| CoCl2·6H2O | 0,0100 g/L | 0,00100 g / 0,000500 g |
| CuSO4·5H2O | 0,0098 g/L | 0,00098 g / 0,000490 g |
| Na2MoO4·2H2O | 0,0063 g/L | 0,00063 g / 0,000315 g |
*(2) F/2 Vitamins solution:
Table 3
| Vitamins solution | Primary stock Conc.[g/L] | Desired stock Conc.[g/L] | g in 100 mL / 50 mL of stock | µL in 100 mL / 50 mL of stock |
|---|---|---|---|---|
| Thyamine·HCl | --- | 0,200 g/L | 0,02 g / 0,01 g | --- |
| Biotin | 0,1 g/L | 0,001 g/L | --- | 1000 µL / 100 µL |
| Cyanocobalamin | 1,0 g/L | 0,001 g/L | --- | 500 µL / 50 µL |
Method:
- Prepare the F/2 trace metal solution (table 2).
Store it in RT.
- Prepare the F/2 vitamins solution (table 3).
Store it in the fridge.
- Prepare the final F/2 medium:
- In 950 mL of filtered artificial/natural seawater, add the quantity of each component (table 1).
NOTE: Don't add the vitamins solution now!
- Bring the final volume to 1 L using filtered artificial/natural seawater.
- Autoclave 20 min at 100ºC.
- Let it cool.
- Add the vitamins solution and filter the medium (0,22 µm).
Under the biological hood.