# 793: MOORELLA GLYCERINI MEDIUM ## Main sol. 793 0.33 g KH2PO4 0.33 g NH4Cl 0.33 g KCl 0.33 g MgCl2 x 6 H2O 0.33 g CaCl2 x 2 H2O 0.5 g Yeast extract 3 ml Glycerol (87%) 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 0.5 ml Sodium resazurin (0.1% w/v) 5 g Na2CO3 1 ml **Wolin's vitamin solution (10x)** 0.5 g Na2S x 9 H2O 1000 ml Distilled water 1. Dissolve ingredients except carbonate, vitamins, and sulfide, then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Dissolve carbonate and adjust pH to 6.7, then dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. ## Trace element solution SL-10 10 ml HCl (25%) 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. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. ## Selenite-tungstate solution 0.5 g NaOH 3 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 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