# J814: TEPIDIMICROBIUM XYLANILYTICUM MEDIUM ## Main sol. J814 ## Solution A 0.49 g KH2PO4 1.19 g Na2HPO4 2 g Tryptone (BD-Difco) 2 g Yeast extract 2 g Peptone 5 g Glucose 0.5 g L-Cysteine HCl x H2O 25 g NaCl 1 mg Resazurin 900 ml Distilled water ## Solution B 0.3 g NH4Cl 0.1 g MgCl2 x 6 H2O 0.01 g CaCl2 x 2 H2O 50 ml Distilled water 1. Autoclave solutions A and B under a N2 atmosphere. After cooling, combine Solutions A and B together with the following solutions (filter-sterilized) under a N2-CO2 (4:1, v/v) gas mixture: 1 ml **Trace element solution** 1 ml **FeCl2 solution** 10 ml **Trace vitamins** 50 ml NaHCO3 (8%) 2. Distribute the medium into culture vessels under the same gas mixture and seal with butyl rubber stoppers. Finally, add per liter the following solution (autoclaved and stored under a N2 atmosphere): 2 ml Na2S x 9 H2O (5%) ## Trace element solution 70 mg ZnCl2 100 mg MnCl2 x 4 H2O 6 mg H3BO3 190 mg CoCl2 x 6 H2O 2 mg CuCl2 x 2 H2O 24 mg NiCl2 x 6 H2O 36 mg Na2MoO4 x 2 H2O 1000 ml Distilled water ## FeCl2 solution 10 ml HCl (7.7 M, 25%) 1.5 g FeCl2 x 4 H2O 990 ml Distilled water ## Trace vitamins 2 mg Biotin 2 mg Folic acid 10 mg Pyridoxine hydrochloride 5 mg Thiamine HCl 5 mg Riboflavin 5 mg Nicotinic acid 5 mg Calcium pantothenate 0.1 mg Vitamin B12 5 mg p-Aminobenzoic acid 5 mg Lipoic acid 1000 ml Distilled water