A+ Medium

A+ Medium Directions For 1 L Total Liquid media: To approximately 900 mL of dH2O add the first 8 components in the order specified while stirring continuously. Bring total volume to 1 L with dH2O. Cover and autoclave medium. Add filter sterilized A+ Trace Components. Allow to cool then store at refrigerator temperature. Agar media: To approximately 390 mL of dH2O add the first 8 components in the order specified while stirring continuously. Bring total volume to 500 mL with dH2O. In a separate container add 15 g of agar to 500 mL of dH2O (Final 1.5% w/v). Cover and autoclave both solutions. In a water bath allow both solutions to cool to 45-50 °C. Add sterile Sodium Thiosulfate to agar solution and mix well. Add filter sterilized A+ Trace Components. Combine both agar and liquid solutions, mix well. Note the agar can solidify quickly. Allow to cool then store at refrigerator temperature. # Component Amount Stock Solution Concentration Final Concentration 1 NaCl(Fisher S271-500) 18 g/L 0.308 M 2 MgSO4•7H2O(Sigma 230391) 5 g/L 0.02 M 3 Na2EDTA•2H2O(Sigma ED255) 10 mL/L 0.3 g/100 mL 0.08 mM 4 KCl(Fisher P 217) 10 mL/L 6 g/100 mL 8.05 mM 5 CaCl2•2H2O(Sigma C-3881) 10 mL/L 3.7 g/100 mL 2.52 mM 6 NaNO3(Fisher BP360-500) 10 mL/L 10 g/100 mL 11.8 mM 7 KH2PO4(Sigma P 0662) 10 mL/L 0.5 g/100 mL 0.37 mM 8 Trizma Base pH 8.2 10 mL/L 10 g/100 mL 8.26 mM 9 A+ Trace Components (sterilize before adding) 10 mL/L 10 Sodium Thiosulfate Pentahydrate (agar media only,sterile)(Baker 3946) 1 mL/L 24.819 g/100 mL 1 µM