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
MES200 mg
DY Trace Metal Solution1.0 ml
f/2 Vitamin Solution0.5 ml
Distilled water to1 L
Adjust pH to 6.8 with NaOH then autoclave after all additions.

DY Trace Metal Solution:
MnCl2200 mg
MnSO440 mg
CoCl28 mg
Na2MoO420 mg
Na3VO42 mg
H2SeO32 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 HCl200.0 mg
Distilled water to1.0 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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