P49 Medium

P49 Medium Suitable for some volvocales, especially Chlamydomonas and Pandorina, which may benefit primarily from the ammonium component, modified volvox medium. Directions For 1 L Total To approximately 900 mL of dH2O, add each of the components in the order specified (except vitamins) while stirring continuously. Adjust the pH to 7.0 Bring total volume to 1 L with dH2O. For 1.5% agar medium add 15 grams 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 MgSO4•7H2O(Sigma 230391) 1 mL/L 4 g/100 mL dH2O 0.16 mM 2 Na2glycerophosphate•5H2O(Sigma G 6501 ) 1 mL/L 5 g/100 mL dH2O 0.16 mM 3 KCl(Fisher P 217) 1 mL/L 5 g/100 mL dH2O 0.67 mM 4 Glycylglycine(Sigma G 1002) 10 mL/L 5 g/100 mL dH2O 3.8 mM 5 P-IV Metal Solution 6 mL/L 6 NH4NO3(Fisher A676) 10 mL/L 1 g/100 mL dH2O 1.2 mM 7 CaCl2•2H2O(Sigma C-3881) 10 mL/L 0.74 g/100 mL dH2O 0.5 mM 8 Yeast extract(Bacto, Difco) 0.2 g/L 9 Tryptone(Sigma T 9410) 0.4 g/L 10 Vitamin B12 1 mL/L 11 Biotin Vitamin Solution 1 mL/L 12 Thiamine Vitamin Solution 1 mL/L