# J1227: ZESTOSPHAERA MEDIUM ## Main sol. J1227 1000 ml **Modified Brock's salt base solution** 2 g Na2S2O3 x 5 H2O 0.16 g NaOH 1 mg Resazurin 10 ml **Trace mineral solution** 1000 ml Distilled water 1. Heat the medium to boiling while bubbling with a N2-CO2 (4:1, v/v) gas mixture. After boiling, cool the medium in a water bath. Continue bubbling with the same gas mixture. While the medium is still warm, add the following: 1 g Yeast extract (Oxoid) 1 g Tryptone 0.5 g Casamino acids (BD-Difco) 0.0625 g L-Cysteine HCl x H2O 2. After the medium is cooled to room temperature, adjust pH to 6.0 - 6.1. Dispense the medium into culture vessels (e.g., 10 ml in Balch type tubes) under the same gas mixture, seal with butyl rubber stoppers and autoclave. ## Modified Brock's salt base solution 1.3 g (NH4)2SO4 0.28 g KH2PO4 0.25 g MgSO4 x 7 H2O 0.07 g CaCl2 x 2 H2O 2 mg FeCl3 x 6 H2O 1.8 mg MnCl2 x 4 H2O 4.5 mg Na2B4O7 x 10 H2O 0.22 mg ZnSO4 x 7 H2O 0.05 mg CuCl2 x 2 H2O 0.03 mg Na2MoO4 x 2 H2O 0.03 mg VOSO4 x n H2O 0.01 mg CoSO4 x 7 H2O 1000 ml Distilled water ## Trace mineral solution 0.5 g Na2-EDTA 0.15 g CoCl2 x 6 H2O 0.1 g MnCl2 x 4 H2O 0.1 g FeSO4 x 7 H2O 0.1 g ZnCl2 0.04 g AlCl3 x 6 H2O 0.03 g Na2WO2 x 2 H2O 0.02 g CuCl2 x 2 H2O 0.02 g NiSO4 x 6 H2O 0.01 g H2SeO3 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 1000 ml Distilled water