# J1136: IRD F1F22 MEDIUM ## Main sol. J1136 0.3 g NH4Cl 0.2 g KH2PO4 0.5 g KCl 0.15 g CaCl2 x 2 H2O 0.4 g MgCl2 x 6 H2O 0.5 g Yeast extract 1 ml **Trace element solution** 0.5 g L-Cysteine HCl x H2O 1 mg Resazurin 960 ml Distilled water 1. Mix components and adjust pH to 7.0. Bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions (autoclaved or *filter-sterilized): 20 ml Glucose (1 M) 20 ml NaHCO3 (*, 5%) 2. Prior to use, add per liter the following solution to reduce the medium (autoclaved and stored under N2): 8 ml Na2S x 9 H2O (5%) ## Trace element solution 12.5 ml HCl (25%) 2.1 g FeSO4 x 7 H2O 30 mg H3BO3 100 mg MnCl2 x 4 H2O 190 mg CoCl2 x 6 H2O 24 mg NiCl2 x 6 H2O 2 mg CuCl2 x 2 H2O 144 mg ZnSO4 x 7 H2O 36 mg Na2MoO4 x 2 H2O 987 ml Distilled water