# J325: MJ/ANHOX-NO3 WITH THIOSULFATE MEDIUM ## Main sol. J325 0.05 g NH4Cl 0.6 g Na2SiO3 0.85 g NaNO3 2.48 g Na2S2O3 x 5 H2O 2 g NaHCO3 1 ml **Trace vitamins** 100 ml **MJ(-N) synthetic seawater** 900 ml Distilled water 1. Distribute medium into culture vessels (e.g., 20 ml of medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers, and autoclave. Adjust pH to 7.5, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v). ## 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 ## MJ(-N) synthetic seawater 30 g NaCl 0.14 g K2HPO4 0.14 g CaCl2 x 2 H2O 3.4 g MgSO4 x 7 H2O 4.18 g MgCl2 x 6 H2O 0.33 g KCl 0.5 mg NiCl2 x 6 H2O 0.5 mg Na2SeO3 x 5 H2O 0.01 g Fe(NH4)2(SO4)2 x 6 H2O 10 ml **Modified Wolin's mineral solution** 1000 ml Distilled water