# 1011b: THERMOSIPHO MJ MEDIUM ## Main sol. 1011b 0.33 g KCl 3.45 g MgSO4 x 7 H2O 2.75 g MgCl2 x 6 H2O 20 g NaCl 0.15 g CaCl2 x 2 H2O 0.25 g NH4Cl 0.15 g K2HPO4 0.01 g Fe(NH4)2(SO4)2 x 6 H2O 0.1 g Yeast extract 1 ml **Trace element solution SL-6** 0.5 ml Sodium resazurin (0.1% w/v) 1 g Na2CO3 2 g D-Glucose 1 ml **Wolin's vitamin solution (10x)** 0.3 g L-Cysteine HCl x H2O 0.3 g Na2S x 9 H2O 1000 ml Distilled water 1. Dissolve ingredients except carbonate, glucose, vitamins, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, glucose, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 6.0 - 6.5, if necessary. ## Trace element solution SL-6 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 ## Wolin's vitamin solution (10x) 20 mg Biotin 20 mg Folic acid 100 mg Pyridoxine hydrochloride 50 mg Thiamine HCl 50 mg Riboflavin 50 mg Nicotinic acid 50 mg Calcium D-(+)-pantothenate 1 mg Vitamin B12 50 mg p-Aminobenzoic acid 50 mg (DL)-alpha-Lipoic acid 1000 ml Distilled water