# 1697: THIOSOCIUS MEDIUM ## Main sol. 1697 16 g NaCl 2.5 g Na2SO4 1 g CaCl2 x 2 H2O 0.45 g KCl 0.25 g NH4Cl 0.14 g K2HPO4 1 ml **Trace elements solution (Pfennig & Lippert,1966)** 20 ml HEPES buffer (1 M, pH 8.0) 6 g MgCl2 x 6 H2O 1 g NaHCO3 2.5 g Na2S2O3 x 5 H2O 1 ml **Wolin's vitamin solution (10x)** 980 ml Distilled water 1. Dissolve ingredients (except HEPES buffer, magnesium chloride, bicarbonate, thiosulfate and vitamins), dispense medium in test tubes or Erlenmeyrer flasks to 20% of volume, close with screw caps and autoclave. Add HEPES buffer, magnesium chloride, bicarbonate, thiosulfate and vitamins to the autoclaved medium from sterile stock solutions sterilized by filtration. Adjust pH of complete medium to 8.0. ## Trace elements solution (Pfennig & Lippert,1966) 5 g EDTA 2.2 g FeSO4 x 7 H2O 0.1 g ZnSO4 x 7 H2O 0.03 g MnCl2 x 4 H2O 0.03 g H3BO3 0.2 g CoCl2 x 6 H2O 0.03 g CuCl2 x 2 H2O 0.03 g NiCl2 x 6 H2O 0.03 g Na2MoO4 x 2 H2O 1000 ml Distilled water 1. pH 3.0-4.0 ## Wolin's vitamin solution (10x) 20 mg Biotin 20 mg Folic acid 100 mg Pyridoxine hydrochloride 50 mg Thiamine HCl 50 mg Riboflavin 50 mg Nicotinic acid 50 mg Calcium D-(+)-pantothenate 1 mg Vitamin B12 50 mg p-Aminobenzoic acid 50 mg (DL)-alpha-Lipoic acid 1000 ml Distilled water