# 1515: OCEANIRHABDUS MEDIUM ## Main sol. 1515 1 g Na2SO4 30 g NaCl 0.7 g KCl 5 g MgCl2 x 6 H2O 1 g CaCl2 x 2 H2O 0.4 g K2HPO4 0.4 g KH2PO4 0.5 g NH4Cl 2 ml Fe(NH4)2(SO4)2 x 6 H2O (0.1% w/v) 1 ml **Trace element solution SL-10** 3 g Yeast extract (OXOID) 4 g Tryptone (BD Bacto) 6.8 g Na-acetate 0.5 ml Sodium resazurin (0.1% w/v) 0.6 g L-Cysteine HCl x H2O 2.5 g NaHCO3 1000 ml Distilled water 1. Dissolve ingredients except cysteine and bicarbonate, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve cysteine, adjust pH to 6.5, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 6.5 - 7.0, if necessary. ## Trace element solution SL-10 10 ml HCl (25%) 1.5 g FeCl2 x 4 H2O 70 mg ZnCl2 100 mg MnCl2 x 4 H2O 6 mg H3BO3 190 mg CoCl2 x 6 H2O 2 mg CuCl2 x 2 H2O 24 mg NiCl2 x 6 H2O 36 mg Na2MoO4 x 2 H2O 990 ml Distilled water 1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.