# 837: DESULFUROMONAS PALMITATIS MEDIUM ## Main sol. 837 880 ml **Solution A** 50 ml **Solution B** 20 ml **Solution C** 1 ml **Solution D** 50 ml **Solution E** 1 ml **Solution F** 1. Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 100% N2 gas atmosphere. Solutions C and F are prepared under 80% N2 and 20% CO2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas atmosphere. Solution C is autoclaved and solutions D, E and F are sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 6.9, if necessary. 2. Note: For transfers use 5 - 10% (v/v) inoculum. ## Solution A 20.8 g NaCl 0.77 g KCl 1 g NH4Cl 0.1 g KH2PO4 0.2 g MgSO4 x 7 H2O 0.02 g CaCl2 x 2 H2O 1.36 g Na-acetate x 3 H2O 10 ml **Modified Wolin's mineral solution** 0.5 ml Sodium resazurin (0.1% w/v) 860 ml Distilled water ## Solution B 10.6 g MgCl2 x 6 H2O 1.5 g CaCl2 x 2 H2O 50 ml Distilled water ## Solution C 1 g Na2CO3 20 ml Distilled water ## Solution D 1 ml **Wolin's vitamin solution (10x)** ## Solution E 8 g Na2-fumarate 50 ml Distilled water ## Solution F 1 ml **Na-dithionite solution (5% w/v)** ## Modified Wolin's mineral solution 1.5 g Nitrilotriacetic acid 3 g MgSO4 x 7 H2O 0.5 g MnSO4 x H2O 1 g NaCl 0.1 g FeSO4 x 7 H2O 0.18 g CoSO4 x 7 H2O 0.1 g CaCl2 x 2 H2O 0.18 g ZnSO4 x 7 H2O 0.01 g CuSO4 x 5 H2O 0.02 g AlK(SO4)2 x 12 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.03 g NiCl2 x 6 H2O 0.3 mg Na2SeO3 x 5 H2O 0.4 mg Na2WO4 x 2 H2O 1000 ml Distilled water 1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. ## 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 ## Na-dithionite solution (5% w/v) 50 g NaHCO3 50 g Na2S2O4 1000 ml Distilled water 1. Dissolve NaHCO3 in water and make the solution anoxic by sparging with 80% N2 and 20% CO2 gas mixture. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in the dark and refrigerated. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.