N/20 Medium

N/20 Medium Directions For 1 L Total To approximately 850 mL of dH2O, add each of the components in the order specified (except vitamins) while stirring continuously. Bring total volume to 1 L with dH2O. Cover and autoclave medium. When cooled, add vitamins. Store at refrigerator temperature. # Component Amount Stock Solution Concentration Final Concentration 1 KH2PO4(Sigma P 0662) 2 mL/L 17.5 g/200 mL dH2O 1.29 mM 2 NaNO3(Fisher BP360-500) 100 µL/L 25 g/200 mL dH2O 0.14 mM 3 K2HPO4(Sigma P 3786) 1 mL/L 15 g/200 mL dH2O 0.43 mM 4 NaCl(Fisher S271-500) 1 mL/L 5 g/200 mL dH2O 0.43 mM 5 CaCl2•2H2O(Sigma C-3881) 1 mL/L 5 g/200 mL dH2O 0.17 mM 6 MgSO4•7H2O(Sigma 230391) 1 mL/L 15 g/200 mL dH2O 0.3 mM 7 P-IV Metal Solution 6 mL/L 8 Sodium acetate(Fisher BP 333) 0.67 g/L 8.12 mM 9 TES buffer(Sigma T 1375) 0.92 g/L 4.01 mM 10 Vitamin B12 1 mL/L 11 Thiamine Vitamin Solution 1 mL/L