# 1166a: DESULFURIBACILLUS MEDIUM (ARSENATE) ## Main sol. 1166a 6 g NaCl 1 g K2HPO4 2.8 g Na2SO4 8 g NaHCO3 22 g Na2CO3 0.2 g NH4Cl 0.2 g MgCl2 x 6 H2O 1 ml **Trace elements solution (Pfennig & Lippert,1966)** 1 ml **Selenite-tungstate solution** 0.05 g Yeast extract 2.2 g Na-pyruvate 6.2 g Na2HAsO4 x 7 H2O 1 ml **Wolin's vitamin solution (10x)** 0.24 g Na2S x 9 H2O 1000 ml Distilled water 1. Dissolve sodium chloride, hydrogenphosphate and sodium sulfate, then sparge solution with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve hydrogencarbonate and carbonate, then dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add ammonium chloride, magnesium chloride, trace elements, yeast extract, pyruvate, arsenate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of vitamins, pyruvate and arsenate should be sterilized by filtration. Adjust pH of complete medium to 9.5 - 10. ## 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 ## Selenite-tungstate solution 0.5 g NaOH 3 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 x 2 H2O 1000 ml Distilled water ## 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