# J843: LIND BHT MEDIUM ## Main sol. J843 0.3 g KH2PO4 0.3 g K2HPO4 1 g NH4Cl 20 g NaCl 0.1 g KCl 0.1 g CaCl2 x 2 H2O 1 ml **Trace element solution** 2 g Yeast extract 4.96 g Na2S2O3 x 5 H2O 0.5 g L-Cysteine HCl x H2O 1 mg Resazurin 925 ml Distilled water 1. Mix components thoroughly and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions from anaerobic stocks (autoclaved or *filter-sterilized): 25 ml NaHCO3 (*, 8%) 30 ml MgCl2 x 6 H2O (10%) 20 ml Sodium acetate (1.0 M) 2. Then, aseptically distribute the medium into culture vessels under a stream of H2-CO2 (4:1, v/v) and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution from an anaerobic stock (autoclave and store under a N2 atmosphere): 8 ml Na2S x 9 H2O (5%) 3. Pressurize inoculated culture vessels to 200 kPa H2-CO2 (4:1, v/v). ## 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