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DY-V Medium

(Andersen et al. 2005, CCMP)

1. Stock solutions

MgSO4•7H2O 5.0 g/100 mL
KCl 0.3 g/100 mL
NH4Cl 0.268 g/100 mL
NaNO3 2.0 g/100 mL
Na2-ß-glycero-phosphate•5H2O 0.216 g/100 mL
H3BO3 0.08 g/100 mL
Na2SiO3•9H2O 1.4 g/100 mL
CaCl2•2H2O 7.5 g/100 mL
FeCl3•6H2O 0.1 g/100 mL
Na2EDTA•2H2O 0.8 g/100 mL

2. Trace metal solutions

MnCl2•4H2O 100 mg
ZnSO4•7H2O (4 g/100 mL) 0.5 mL
CoCl2•6H2O (0.8 g/100 mL) 0.5 mL
Na2MoO4•2H2O (1.19 g/100 mL) 0.5 mL
Na3VO4•nH2O (0.2 g/100 mL) 0.5 mL
H2SeO3 (0.4 g/100 mL) 0.5 mL
dH2O to 500 mL

3. 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.

4. 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 to 100 mL.
The vitamin working stock solution is divided into to 10-mL lots in polyethylene vials and kept frozen until use.

5. Final preparation of the DY-V medium
To 950 mL dH2O add 200 mg MES buffer, 1 mL of each of the stock solution, 1 ml of the trace metal solution and 0.5 mL f/2 vitamin solution. Make final volume up to 1 L with dH2O. Adjust pH to 6.8 with NaOH, then autoclave.

Andersen, R.A., Berges, J.A., Harrison, P.J. & Watanabe, M.M. 1997. Recipes for freshwater and seawater media.
In: R. A. Andersen ed.: Algal culturing techniques. Elsevier Academic Press, Burlington, p. 429-538. Link