# 943: SEDIMENTIBACTER SAALENSIS MEDIUM ## Main sol. 943 0.45 g (NH4)HCO3 0.12 g CaCl2 x 2 H2O 0.13 g MgSO4 x 6 H2O 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 3 g Yeast extract 3.8 g NaHCO3 0.5 ml Sodium resazurin (0.1% w/v) 10 ml Sodium Potassium phosphate buffer (1 M, pH 7.3) 0.75 g Glycine 2.11 g L-Arginine x HCl 1 ml **Seven vitamins solution** 0.15 g Na2S x 9 H2O (3% w/v) 1000 ml Distilled water 1. Dissolve ingredients (except bicarbonate, phosphate buffer, glycine, arginine, vitamins, and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and adjust pH to 7.3, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add phosphate buffer, glycine, arginine, vitamins (sterilized by filtration), and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Prior to inoculation add 10 - 20 mg sodium dithionite per liter medium from a 5% (w/v) stock solution, freshly prepared under 100% N2 gas and filter-sterilized. ## 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 ## Seven vitamins solution 100 mg Vitamin B12 80 mg p-Aminobenzoic acid 20 mg D-(+)-biotin 200 mg Nicotinic acid 100 mg Calcium pantothenate 300 mg Pyridoxine hydrochloride 200 mg Thiamine-HCl x 2 H2O 1000 ml Distilled water