# J536: PHE91 MEDIUM ## Main sol. J536 969 ml **Basal solution** 1 ml **SL-7 trace element solution** 1 ml **Vitamin solution** 1 ml **Vitamin B12 solution 0.05 g/L** 1 ml **Selenite-tungstate solution** 1 ml CaCl2 x 2 H2O (68 mM) 10 ml K2HPO4 (609 mM) 1 ml Resazurin (0.02%) 5 ml Dithionite (0.2 M) 10 ml Pyruvate (1 M) 1. Autoclave the basal solution under a N2 gas atmosphere. Separately filter-sterilize the other solutions and add to the medium anaerobically and aseptically. Prepare dithionite solution just before use. ## Basal solution 0.46 g NH4H2PO4 0.49 g MgSO4 x 7 H2O 969 ml Distilled water ## SL-7 trace element solution 10 ml HCl (7.7 M, 25%) 1.5 g FeCl2 x 4 H2O 190 mg CoCl2 x 6 H2O 100 mg MnCl2 x 4 H2O 70 mg ZnCl2 62 mg H3BO3 36 mg Na2MoO4 x 2 H2O 24 mg NiCl2 x 6 H2O 17 mg CuCl2 x 2 H2O 1000 ml Distilled water 1. Initially, dissolve FeCl2 x 4H2O in 25% HCl, then add distilled water followed by the other components. ## Vitamin solution 2.5 mg Biotin 25 mg Nicotinic acid 12.5 mg Thiamine HCl 12.5 mg p-Aminobenzoic acid 6.5 mg DL-Calcium pantothenate 62.5 mg Pyridoxine hydrochloride 1000 ml Distilled water 1. Store in the dark at 4C. ## Vitamin B12 solution 0.05 g/L 5 mg Vitamin B12 100 ml Distilled water ## Selenite-tungstate solution 0.4 g NaOH 6 mg Na2SeO3 x 5 H2O 8 mg Na2WO4 x 2 H2O 1000 ml Distilled water