# 1475: METALLIBACTERIUM MEDIUM ## Main sol. 1475 132 mg (NH4)2SO4 53 mg MgCl2 x 6 H2O 27 mg KH2PO4 147 mg CaCl2 x 2 H2O 0.5 mg VOSO4 x 5 H2O 1 ml **Wolfe's mineral elixir** 10 mg FeSO4 x 7 H2O 1.4 g Trypticase peptone (BD BBL) 1 ml **Wolin's vitamin solution (10x)** 1000 ml Distilled water 1. Dissolve ingredients except vitamins and ferrous sulfate and adjust pH of solution to 5.0. Dissolve ferrous sulfate (1% w/v) in 0.1 N H2SO4 and add to the medium prior to autoclaving. After autoclaving add vitamins from a stock solution that has been sterilized by filtration. ## Wolfe's mineral elixir 30 g MgSO4 x 7 H2O 5 g MnSO4 x H2O 10 g NaCl 1 g FeSO4 x 7 H2O 1.8 g CoCl2 x 6 H2O 1 g CaCl2 x 2 H2O 1.8 g ZnSO4 x 7 H2O 0.1 g CuSO4 x 5 H2O 0.18 g AlK(SO4)2 x 12 H2O 0.1 g H3BO3 0.1 g Na2MoO4 x 2 H2O 2.8 g (NH4)2Ni(SO4)2 x 6 H2O 0.1 g Na2WO4 x 2 H2O 0.1 g Na2SeO4 1000 ml Distilled water 1. First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts. ## 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