# 764: HALOANAEROBIUM LACUSROSEUS MEDIUM ## Main sol. 764 1 g NH4Cl 0.3 g K2HPO4 0.3 g KH2PO4 2 g MgCl2 x 6 H2O 0.2 g CaCl2 x 2 H2O 4 g KCl 1 g Na-acetate 10 g Glucose 0.5 g Trypticase 200 g NaCl 1 ml **Modified Wolin's mineral solution II** 1 mg Resazurin 1 g Yeast extract 1000 ml Distilled water 1. Prepare the medium and boil, cool under N2:CO2 (80:20 v/v). Dispense medium into Hungate tubes or serum bottles under N2:CO2 (80:20 v/v), autoclave. To the sterile medium add from sterile stock solutions 2% Na2S x 9 H2O (0.1ml/10ml), 10% NaHCO3 (0.5ml/10ml), and 1M glucose (0.25ml/10ml). Final pH 7.0. ## Modified Wolin's mineral solution II 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 0.025 g NiCl2 x 6 H2O 0.3 mg Na2SeO3 x 5 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.