# 465a: MINERAL MEDIUM WITH 2-HYDROXYBIPHENYL ## Main sol. 465a 1000 ml **Main sol. 465** 0.5 g/l [1,1′-Biphenyl]-2-ol (dissolved in 50 g/l ethanol) 1. Prepare medium 465. Dissolve 2-hydroxybiphenyl in ethanol (50 g/l) and filter sterilize. Pipette a volume of this stock solution into a sterile culture vessel so that the final concentration of 2-hydroxybiphenyl will be 0.5 g/l medium. Let the ethanol evaporate under sterile conditions. Add medium 465. ## Main sol. 465 3.5 g Na2HPO4 x 2 H2O 1 g KH2PO4 0.5 g (NH4)2SO4 0.1 g MgCl2 x 6 H2O 0.05 g Ca(NO3)2 x 4 H2O 1 ml **Trace element solution SL-4** 1000 ml Distilled water 1. pH 7.25 2. Rehydrate and cultivate lyophilized cells in the complex medium recommended for the specific strain (e.g. medium 1, 220, 535 or 830). After this reactivation, cultivate on mineral medium with the appropriate carbon source. ## Trace element solution SL-4 0.5 g Na2-EDTA 0.2 g FeSO4 x 7 H2O 0.1 g ZnSO4 x 7 H2O 0.03 g MnCl2 x 4 H2O 0.3 g H3BO3 0.2 g CoCl2 x 6 H2O 0.01 g CuCl2 x 2 H2O 0.02 g NiCl2 x 6 H2O 0.03 g Na2MoO4 x 2 H2O 1000 ml Distilled water 1. First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. ## Main sol. 1 5 g Peptone 3 g Meat extract 15 g Agar, for solid medium 1000 ml Distilled water 1. Adjust pH to 7.0. 2. For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation. ## Main sol. 220 15 g Casein peptone 5 g Soy peptone 5 g NaCl 15 g Agar 1000 ml Distilled water 1. Adjust pH to 7.3. Medium is identical with Tryptone Soya Agar (Oxoid CM 131). ## Main sol. 535 30 g Trypticase soy broth (BBL 11768, Oxoid CM129 or Merck 5459) 20 g Agar 1000 ml Distilled water 1. pH 7.3 2. Autoclave at 121°C for 15 min. ## Main sol. 830 0.5 g Yeast extract 0.5 g Proteose peptone (Difco no. 3) 0.5 g Casamino acids 0.5 g Glucose 0.5 g Starch (soluble) 0.3 g Na-pyruvate 0.3 g K2HPO4 0.05 g MgSO4 x 7 H2O 15 g Agar 1000 ml Distilled water 1. Final pH 7.2; adjust with crystalline K2HPO4 or KH2PO4 before adding agar. Add agar, heat medium to boiling to dissolve agar, and autoclave for 15 min at 121°C.