Modified Bold 3N Medium

Modified Bold 3N Medium Modification of Bold's recipe. General purpose freshwater medium used for xenic cultures, especially blue-greens and reds. Directions For 1 L Total pH 6.2 To approximately 850 mL of dH2O, add each of the components in the order specified (except vitamins) while stirring continuously. Bring the total volume to 1 L with dH2O. For 1.5% agar medium add 15 g of agar into the flask; do not mix. Cover and autoclave medium. When cooled add vitamins. For agar medium add vitamins, mix, and dispense before agar solidifies. Store at refrigerator temperature. # Component Amount Stock Solution Concentration Final Concentration 1 NaNO3(Fisher BP360-500) 30 mL/L 10 g/400mL dH2O 8.82 mM 2 CaCl2•2H2O(Sigma C-3881) 10 mL/L 1 g/400mL dH2O 0.17 mM 3 MgSO4•7H2O(Sigma 230391) 10 mL/L 3 g/400mL dH2O 0.3 mM 4 K2HPO4(Sigma P 3786) 10 mL/L 3 g/400mL dH2O 0.43 mM 5 KH2PO4(Sigma P 0662) 10 mL/L 7 g/400mL dH2O 1.29 mM 6 NaCl(Fisher S271-500) 10 mL/L 1 g/400mL dH2O 0.43 mM 7 P-IV Metal Solution 6 mL/L 8 Soilwater: GR+ Medium 40 mL/L 9 Vitamin B12 1 mL/L 10 Biotin Vitamin Solution 1 mL/L 11 Thiamine Vitamin Solution 1 mL/L