KH2PO4 0.30 g Na2HPO4 x 12 H2O 5.30 g NH4Cl 1.00 g MgCl2 x 6 H2O 0.20 g Trace element solution 10.00 ml FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4) 1.50 ml Yeast extract 1.00 g Sodium resazurin (0.1% w/v) 0.50 ml D-Glucose 5.00 g Wolin's vitamin solution 5.00 ml Na2S x 9 H2O 0.50 g Distilled water 1000.00 ml Dissolve ingredients (except glucose, vitamins and sulfide), adjust pH to 6.0, sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense under same gas atmosphere into Hungate-type tubes or serum vials and autoclave. After sterilization add glucose and sulfide from anoxic stock solutions autoclaved under 100% N2 gas and vitamins from a filter-sterilized anoxic stock solution prepared under 100% N2. Adjust pH of complete medium to 6.0 - 6.5, if necessary. For DSM 8683, DSM 8684, DSM 16487: Adjust pH of complete medium to 5.5. Trace element solution (from medium 144) Nitrilotriacetic acid (NTA) 12.80 g FeCl2 x 4 H2O 0.20 g MnCl2 x 4 H2O 0.10 g CoCl2 x 6 H2O 0.17 g CaCl2 x 2 H2O 0.10 g ZnCl2 0.10 g CuCl2 0.02 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.01 g NiCl2 x 6 H2O 0.03 g NaCl 1.00 g Na2SeO3 x 5 H2O 0.03 g Distilled water 1000.00 ml First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml. Wolin's vitamin solution (from medium 141) Biotin 2.00 mg Folic acid 2.00 mg Pyridoxine hydrochloride 10.00 mg Thiamine HCl 5.00 mg Riboflavin 5.00 mg Nicotinic acid 5.00 mg Calcium D-(+)-pantothenate 5.00 mg Vitamin B12 0.10 mg p-Aminobenzoic acid 5.00 mg (DL)-alpha-Lipoic acid 5.00 mg Distilled water 1000.00 ml
