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L16, L16-V

(Lindström 1991)

This medium was originally formulated for growth of Peridinium gatunense from Lake Kinneret (Lindström 1991), but works very well for other dinoflagellates as well. The conductivity is c. 140 µS·cm-1, but c. 280 µS·cm-1 and c. 560 µS·cm-1 in 2xL16 and 4xL16 versions, respectively. Originally, no vitamins were added but we routinely add f/2 vitamins.

1. Stock solutions

NaHCO3 0.840 g/100 mL
NaNO3 1.280 g/100 mL
CaCl2•2H2O 1.470 g/100 mL
MgSO4•7H2O 1.230 g/100 mL
Na2O3Si•9H2O 1.140 g/100 mL
K2HPO4 0.174 g/100 mL

2. Trace element solution

Na2EDTA•2H2O 0.558 g
FeCl3•6H2O 0.108 g
MnCl2•4H2O 0.010 g
ZnCl2 0.027 g
CoCl2•6H2O 0.0096 g
Na2MoO4•2H2O 0.048 g
dH2O to 1000 mL

3. Selenium-solution

Na2SeO3* 0.00129 g
dH2O to 1000 mL

* Na2SeO3•5H2O 0.00184g/L is used in original recipe.

4. Preparation of L16-medium
Add 10 mL of stock 1.1 mL of stock solutions 2-5, 1 mL trace element solution and 100 µL selenium solution to 900 mL dH2O. Fill up to 1000 mL and autoclave.

For 2xL16 and 4xL16 add 2 or 4 times of every constituent, respectively.
However, add stock solution 6 aseptically after autoclaving and cooling in order to avoid precipitation.

Add 0.5 mL f/2 vitamins before autoclaving.
Notice! We do not increase the vitamin concentration in 2xL16-V and 4xL16-V.

5. f/2 vitamin primary stock solutions

Biotin 0.01g/100 mL
Cyanocobalamin (B12) 0.1g/100 mL

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).
Keep the vitamin solutions frozen. Bottles of polyethylene are recommended for storage of vitamins.

6. f/2 vitamin working stock solution
1) Dissolve 20 mg Thiamine HCl (Vitamin B1) in ca. 80 mL dH2O in a 100 mL volumetric flask.
2) Add 1 mL of the biotin primary stock solution.
3) Add 0.1 mL of the cyanocobalamin primary stock solution.
4) Fill with dH2O to100 mL.
The vitamin working stock solution is divided into to 10-mL lots in polyethylene vials and kept frozen until use.

Lindström, K. 1991. Nutrient requirements of the dinoflagellate Peridinium gatunense. J. Phycol. 27: 207-219.