# J671: THERMOANAEROBIUM MEDIUM ## Main sol. J671 3 g Yeast extract 10 g Tryptone (BD-Difco) 0.9 g NH4Cl 0.9 g NaCl 0.4 g MgCl2 x 6 H2O 0.75 g KH2PO4 1.5 g K2HPO4 9 ml **Trace element solution** 5 ml **Trace vitamins** 3 mg FeSO4 x 7 H2O 1 mg Resazurin 950 ml Distilled water 1. Mix ingredients and adjust pH to 7.0. Distribute the medium into culture vessels under a N2 atmosphere, seal with butyl rubber stoppers and autoclave. Separately autoclave the following solutions under a N2 atmosphere and add to the medium (per 950 ml): 50 ml Glucose (10%) 10 ml Na2S x 9 H2O (5%) 2. Readjust pH of the medium to 7.2 - 7.4 with sterile NaOH, if necessary. ## Trace element solution 12.8 g Nitrilotriacetic acid 0.2 g FeCl2 x 4 H2O 0.1 g MnCl2 x 4 H2O 0.17 g CoCl2 x 6 H2O 0.1 g CaCl2 x 2 H2O 0.1 g ZnCl2 0.02 g CuCl2 x 2 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.026 g NiCl2 x 6 H2O 1 g NaCl 0.02 g Na2SeO3 1000 ml Distilled water 1. Dissolve nitrilotriacetic acid first in 200 ml of distilled water, and adjust pH 6.5 with KOH. Then add the remaining salts and adjust volume to 1000 ml. ## Trace vitamins 2 mg Biotin 2 mg Folic acid 10 mg Pyridoxine hydrochloride 5 mg Thiamine HCl 5 mg Riboflavin 5 mg Nicotinic acid 5 mg Calcium pantothenate 0.1 mg Vitamin B12 5 mg p-Aminobenzoic acid 5 mg Lipoic acid 1000 ml Distilled water