L1 Medium and Derivatives

NaNO3 (75.0 g/L dH2O) 1.0 ml
NaH2PO4 H2O (5.0 g/L dH2O) 1.0 ml
Na2SiO3 9H2O (30.0 g/L dH2O) 1.0 ml
L1 Trace Metal Solution 1.0 ml
f/2 Vitamin Solution 0.5 ml
Filtered seawater to 1.0 L

Autoclave after all additions.

L1 Trace Metal Solution:
FeCl3 6H2O 3.15 g
Na2EDTA 2H2O 4.36 g
CuSO4 5H2O (9.8 g/L dH2O) 0.25 ml
Na2MoO4 2H2O (6.3 g/L dH2O) 3.0 ml
ZnSO4 7H2O (22.0 g/L dH2O) 1.0 ml
CoCl2 6H2O (10.0 g/L dH2O) 1.0 ml
MnCl2 4H2O (180.0 g/L dH2O) 1.0 ml
H2SeO3 (1.3 g/L dH2O) 1.0 ml
NiSO4 6H2O (2.7 g/L dH2O) 1.0 ml
Na3VO4 (1.84 g/L dH2O) 1.0 ml
K2CrO4 (1.94 g/L dH2O) 1.0 ml
Filtered seawater to 1.0 L

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
Filter sterilize into plastic vials and store in refrigerator.

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).

L1 Derivatives

L1-Si:
Prepare as for L1 Medium but omit Na2SiO3 9H2O.

L1-Si (80%SW):
This medium is used to maintain organisms from brackish waters (approximately 26 ppt salinity). Mix 800 ml filtered seawater to 200 ml dH2O. Prepare L1-Si Medium in the diluted seawater, then autoclave.

L2 Trace Metal Solution:
This trace metal solution is similar to L1 Trace Metal Solution, but contains raised levels of Na2EDTA 2H2O. The higher chelation is favoured by some oceanic strains. Prepare using the recipe for L1 Trace Metal Solution, but add a total of 37.2 g of Na2EDTA 2H2O.


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