Freshwater Trace Metal-Buffered Medium
CaCl2 | 1 mL |
MgSO4 | 1 mL |
NaHCO3 | 1 mL |
K2HPO4 | 1 mL |
NaNO3 | 1 mL |
Trace metal mix | 1 mL |
f/2 vitamin mix | 1 mL |
Milli-Q Water | 993 mL |
Individual Salt Stock Solutions:
CaCl2·2 H2O | 18.4 g |
MgSO4·7 H2O | 18.5 g |
NaHCO3 | 6.3 g |
K2HPO4 | 0.87 g |
NaNO3 | 4.25 g |
Na2SiO3·9 H2O | 1.775g |
Individual Trace Metals Stocks Solutions:
CuSO4 · 5 H2O | 0.125 g | 500ml |
(NH4)6Mo7O24 · 4 H2O | 0.133 g | 500 mL |
CoCl2 · 6 H2O | 0.298 g | 500 mL |
MnCl2 · 4 H2O | 0.455 g | 100 mL |
ZnSO4 · 7 H2O | 0.115 g | 100 mL |
FeCl3 · 7 H2O | 1.352 g | 500 mL |
Trace Metals Mix
To create different media having known Fe concetration:
1 microM Fe (final) stock
milli-Q H2O | 443 mL |
CuSO4·5 H2O | 500 microL |
(NH4)6Mo7O24·4 H2O | 500 microL |
CoCl2·6 H2O | 500 microL |
MnCl2·4 H2O | 500 microL |
ZnSO4·7 H2O | 500 microL |
Na2EDTA (0.5 M) | 5 mL |
FeCl3·7 H2O | 50 mL |
100 nM Fe (final) stock
milli-Q H2O | 448 mL |
CuSO4·5 H2O | 500 microL |
(NH4)6Mo7O24·4 H2O | 500 microL |
CoCl2·6 H2O | 500 microL |
MnCl2·4 H2O | 500 microL |
ZnSO4·7 H2O | 500 microL |
Na2EDTA (0.5 M) | 5 mL |
FeCl3·7 H2O | 5 mL |
10 nM Fe (final) stock
milli-Q H2O | 492 mL |
CuSO4·5 H2O | 500 microL |
(NH4)6Mo7O24·4 H2O | 500 microL |
CoCl2·6 H2O | 500 microL |
MnCl2·4 H2O | 500 microL |
ZnSO4·7 H2O | 500 microL |
Na2EDTA (0.5 M) | 5 mL |
FeCl3·7 H2O | 500 microL |
Add the indicated volume of milli-Q H2O to each acid-clean polyethylene bottle and then add the indicated amount of each trace metal stock. Agitate manually following each addition. Ensure the solution is particularly well-mixed following addition of EDTA and prior to addition of Fe.
f/2 Vitamin Solution:
Vitamin B12 (1.0 g/L dH2O) | 1.0 ml |
Biotin (0.1 g/L dH2O) | 10.0 ml |
Thiamine HCl | 200.0 mg |
Distilled water to | 1.0 L |
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 1.0 mg of Vitamin B12 add 0.89 ml dH2O). When preparing the Biotin Stock Solution allow for approximately 4% water of crystallization (For each 1.0 mg of Biotin add 9.6 ml dH2O).
Morel et al. 1975
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