|
Stocks |
|
Phosphate Stock Solution: |
|
| NaH2PO4 H2O |
1 g |
In 200 mL Distilled H20
|
|
|
Trace Metal Stock Solution: |
|
| FeCl3 |
0.13 g |
| EDTA |
0.87 g |
| CuSO4 5H2O 0.98 g/100 mL |
100 microL |
| ZnSO4 7H2O 2,2 g /100 mL |
100 microL |
| CoCl2 6H2O 1 g/100 mL |
100 microL |
| MnCl2 4H2O 18 g/100 mL |
100 microL |
| Na2MoO4 2H2O 0.63 g/100 mL |
100 microL |
| In 200 mL Distilled H20 |
|
|
Vitamin Stock: |
|
| Biotin |
250 microL |
| Vitamin B12 |
250 microL |
| Thiamine HCl |
50.0 mg |
| In 250 m L Distilled H20 |
Aliquot of 200 microL (or 400 microL) in 1.5 mL tubes
Freeze and store at -20 C |
|
|
NaHCO3 (Fresh Solution): |
|
| NaHCO3 |
0.42 g |
| To 10 L Distilled H20 |
| Filter-sterilise before adding to F/2 solution |
|
|
Agar Plates |
|
- Prepare medium with 2X stock solution (no vitamins), but same amount of Artificial Sea Water (typically 200 mL) + (optionally add 20 mM HEPES from filter-sterilised solution)
- Prepare 3% w/v Noble Agar solution in double distilled water , and autoclave (typically 200 mL in a 500 mL bottle)
- Let the autoclaved agar solution cool to about 40 C (in a water bath),
- Add the 2X F/2 solution, mix, add vitamins and pour plates
|
|
F/2 + Artificial Seawater recipeit in PDF format: F2.pdf |
