F2培养基配制.docVIP

f/2 Medium (Guillard Ryther 1962, Guillard 1975) To 950 mL filtered seawater add: Quantity Compound Stock Solution Molar Concentration in Final Medium 1 mL NaNO3 75 g/L dH2O 8.83 x 10-4 M 1 mL NaH2PO4 · H2O 5 g/L dH2O 3.63 x 10-5 M 1 mL * Na2SiO3 · 9H2O* 30 g/L dH2O* 1.07 x 10-4 M* 1 mL f/2 trace metal solution (see recipe below) - 0.5 mL f/2 vitamin solution (see recipe below) - Make final volume up to 1 L with filtered seawater and autoclave. *Note: Autoclaved f/2 medium produces extensive silica precipitate. We delete silicate when it is not required by the alga (see f/2-Si medium below). ? f/2 Trace Metal Solution(Guillard Ryther 1962, Guillard 1975) To 950 mL dH2O add: Quantity Compound Stock Solution Molar Concentration in Final Medium 3.15 g FeCl3 · 6H2O - 1 x 10-5 M 4.36 g Na2EDTA · 2H2O - 1 x 10-5 M 1 mL CuSO4 · 5H2O 9.8 g/L dH2O 4 x 10-8 M 1 mL Na2MoO4 · 2H2O 6.3 g/L dH2O 3 x 10-8 M 1 mL ZnSO4 · 7H2O 22.0 g/L dH2O 8 x 10-8 M 1 mL CoCl2 · 6H2O 10.0 g/L dH2O 5 x 10-8 M 1 mL MnCl2 · 4H2O 180.0 g/L dH2O 9 x 10-7 M Make final volume up to 1 L with dH2O. Autoclave. ? f/2 Vitamin Solution(Guillard Ryther 1962, Guillard 1975) To 950 mL dH2O add: Quantity Compound Stock Solution Molar Concentration in Final Medium 1 mL Vitamin B12 (cyanocobalamin) 1.0 g/L dH2O 1 x 10-10 M 10 mL Biotin 0.1 g/L dH2O 2 x 10-9 M 200 mg Thiamine · HCl - 3 x 10-7 M Make final volume up to 1 L with dH2O. Autoclave 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 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 mg of biotin, add 9.6 mL dH2O). ? f/2 Derivatives Black Sea Medium: For brackish water organisms (16 psu, half-strength nutrients). Combine 500 mL f/2 medium and 500 mL dH2O. Autoclave.