Adapted from (Harrison et al. 1980, Berges et al. 2001)
1 ENRICHED SEAWATER MEDIUM RECIPE
Either natural (ESNW) or artificial (ESAW) seawater may be used. See section 1.1 for artificial seawater recipe. If using natural seawater and autoclaving the solution, 0.12 g of sodium bicarbonate can be added per liter of seawater to minimize precipitation.
Add the indicated amounts of each stock solution to 950 ml of seawater. Bring final volume to 1 liter with seawater. The final pH should be 8.2. Sterilize medium with 0.22 μm pore size Gelman filter.
1.1 ARTIFICIAL SEAWATER RECIPE
Dissolve the anhydrous salts in 600 mL of dH2O. and dissolve the hydrated salts in 300 mL dH2O. Combine salt solutions I and II. Bring the final volume to 1 liter with dH2O. Autoclaving the final medium often causes precipitates to form. If autoclaving is necessary, autoclave the two salt solutions separately, and when they are completely cooled, aseptically combine them.
1.1.1 Salt solution I: Anhydrous salts
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
NaCl
21.19
3.63 x 10-1
Na2SO4
3.55
2.50 x 10-2
KCl
0.599
8.04 x 10-3
NaHCO3
0.174
2.07 x 10-3
KBr
0.0863
7.25 x 10-4
H3BO3
0.0230
3.72 x 10-4
NaF
0.0028
6.57 x 10-5
1.1.2 Salt solution II: Hydrated salts
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
MgCl2⋅6H2O
9.592
4.71 x 10-2
CaCl2⋅2H2O
1.344
9.14 x 10-3
SrCl2⋅6H2O
0.0218
8.18 x 10-5
1.2 NUTRIENT STOCK RECIPES
1.2.1 Enrichment stock I: Nitrate
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
NaNO3
46.7
5.49 x 10-4
1.2.2 Enrichment stock II: Phosphate
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
NaH2 PO4⋅H2O
3.09
2.1 x 10-5
1.2.3 Enrichment stock III: Iron
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
Na2EDTA⋅2H2O
3.09
6.56 x 10-6
FeCl2⋅6H2O
1.77
6.55 x 10-6
1.2.4 Enrichment stock IV: Borate
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
H3BO3
3.80
6.146 x 10-5
1.2.5 Enrichment stock V: Silica
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
Na2SiO3⋅9H2O
15.00
1.05 x 10-4
1.2.6 Trace metals stock solution
Component
Stock Concentration (g⋅L-1)
Concentration in final medium (M)
Na2EDTA⋅2H2O
2.44
8.29 x 10-6
ZnSO4⋅7H2O
0.073
2.54 x 10-7
CoSO4⋅7H2O
0.016
5.69 x 10-8
MnSO4⋅4H2O
0.54
2.42 x 10-6
Na2MoO4⋅2H2O
1.48 x 10-3
6.12 x 10-9
Na2SeO3
1.73 x 10-4
1.00 x 10-9
NiCl6⋅6H2O
1.49 x 10-3
6.3 x 10-9
1.3 VITAMIN STOCK RECIPE
Into 900 mL dH2O add the components listed below and bring the final volume to 1 liter with dH2O. Filter-sterilize and store frozen.
