Provasoli Solution
Solution 1 (10×)
| Reagent | Quantity (for 1 L) | Final concentration |
|---|---|---|
| H3BO3 (MW = 61.83) | 1.9 g | 30.7 mm |
| FeCl3 (MW = 162.21) | 0.05 g | 0.3 mm |
| MnSO4·H2O (MW = 169.02) | 0.273 g | 1.6 mm |
| ZnSO4·7H2O (MW = 287.54) | 0.0367 g | 0.127 mm |
| CoSO4·7H2O (MW = 281.1) | 0.008 g | 28 µm |
| EDTA (0.5 m, pH 8) | 11.4 mL | 5.7 mm |
Solution 2 (10×)
| Reagent | Quantity (for 500 mL) |
|---|---|
| Vitamin B12 (cyanocobalamine) | 3.35 mg |
| Thiamine hydrochloride (vitamin B1) (MW = 337.27) | 165 mg |
| Biotin C10H16N2O3S (MW = 244.31) | 1.65 mg |
| TRIS (Trisma base) C4H11NO3 (MW = 121.14) | 166.5 g |
Solution 3 (10×)
| Reagent | Quantity (for 1 L) | Final concentration |
|---|---|---|
| (NH4)2Fe(SO4)2·6H2O (MW = 392.14) | 1.17 g | 3 mm |
| EDTA (0.5 m, pH 8) | 6.8 mL | 3.4 mm |
Solution 4 (10×)
| Reagent | Quantity (for 1 L) | Final concentration |
|---|---|---|
| NaNO3 (MW = 84.99) | 23 g | 270 mm |
Solution 5 (10×)
| Reagent | Quantity (for 1 L) | Final concentration |
|---|---|---|
| C3H7Na2O6P·5H2O “glycerophosphate” (MW = 216.04) | 3.33 g | 15.4 mm |
Prepare each stock solution separately, autoclave, and store in glass bottles at 4°C. Use a dark bottle for Solution 2.
For 1 L of Provasoli solution, add 100 mL of each of Solutions 1, 3, 4, and 5 plus 10 mL of Solution 2 to MilliQ water (the starting pH should be between 9.6 and 9.8). Adjust to pH 7.8 with concentrated HCl (37%) and adjust the volume to 1 L with MilliQ water. Aliquot into small glass bottles (20, 50, 100, or 200 mL), autoclave, and store at 4°C.
- © 2012 Cold Spring Harbor Laboratory Press
