# 733: SULFITOBACTER PONTIACUS MEDIUM ## Main sol. 733 20 mM Na-acetate 8 g HEPES 1 g K2HPO4 0.5 g NH4Cl 1 g Yeast extract 0.5 g Peptone 15 g NaCl 1 g MgSO4 x 7 H2O 0.05 g CaCl2 x 2 H2O 1 ml **Modified Wolin's mineral solution** 0.1 g Biotin 1000 ml Distilled water 1. Prepare the HEPES, NaCl, K2HPO4 and NH4Cl as a basal salt solution, adjusting the pH to 7.5-7.8 with NaOH before sterilisation. All other components are added from sterile stock solutions. ## Modified Wolin's mineral solution 1.5 g Nitrilotriacetic acid 3 g MgSO4 x 7 H2O 0.5 g MnSO4 x H2O 1 g NaCl 0.1 g FeSO4 x 7 H2O 0.18 g CoSO4 x 7 H2O 0.1 g CaCl2 x 2 H2O 0.18 g ZnSO4 x 7 H2O 0.01 g CuSO4 x 5 H2O 0.02 g AlK(SO4)2 x 12 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.03 g NiCl2 x 6 H2O 0.3 mg Na2SeO3 x 5 H2O 0.4 mg Na2WO4 x 2 H2O 1000 ml Distilled water 1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.