DYIV Medium

KCl (3.0 g/L dH2O) 1.0 ml
NH4Cl (2.68 g/L dH2O) 1.0 ml
NaNO3 (20.0 g/L dH20) 1.0 ml
beta-Glycerophosphate (2.16 g/L dH2O) 1.0 ml
H3BO3 (0.8 g/L dH2O) 1.0 ml
Na2EDTA 2H2O (8.0 g/L dH2O) 1.0 ml
Na2SiO3 9H2O (14.0 g/L dH2O) 1.0 ml
FeCl3 (1.0 g/L dH2O) 1.0 ml
CaCl2 (75.0 g/L dH2O) 1.0 ml
MES 200 mg
DY Trace Metal Solution 1.0 ml
f/2 Vitamin Solution 0.5 ml
Distilled water to 1 L

Adjust pH to 6.8 with NaOH then autoclave after all additions.

DY Trace Metal Solution:
MnCl2 200 mg
MnSO4 40 mg
CoCl2 8 mg
Na2MoO4 20 mg
Na3VO4 2 mg
H2SeO3 2 mg

Dissolve the metal compounds individually (each in 100 mL dH2O). Mix the six solutions then bring volume up to 1.0 L with dH2O.

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).

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