# J1312: MODIFIED MV MEDIUM ## Main sol. J1312 1. Mix ingredients, bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream and autoclave under the same gas mixture. After cooling, aseptically and anaerobically add per liter the following solutions (filter-sterilized): 1 g NH4Cl 2.1 g NaCl 0.825 g MgCl2 x 6 H2O 0.1 g CaCl2 x 2 H2O 0.5 g KH2PO4 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 1 g Yeast extract (BD-Difco) 0.5 ml Resazurin (0.1%) 10 ml **Trace vitamins** 30 ml NaHCO3 (8%) 20 ml Taurine (0.5 M) 20 ml Sodium pyruvate (1 M) 20 ml Sodium L-lactate (0.5 M) 10 ml **Hemin solution** 2. Prior to inoculation, aseptically and anaerobically add 0.01 volume of L-cysteine x HCl x H2O to the medium (3% solution, autoclaved and stored under a N2 atmosphere). ## Trace element solution SL-10 10 ml HCl (25%, 7.7 M) 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. Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. ## Selenite-tungstate solution 0.5 g NaOH 3 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 x 2 H2O 1000 ml Distilled water ## 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