Final pH: 9-.8 10.0 Final volume: 1000 ml MJ(-N) synthetic seawater 100.00 ml Yeast extract (BD Bacto) 2.00 g Tryptone (BD Bacto) 2.00 g Na2-tartrate x 2 H2O 1.00 g Na3-citrate 1.00 g NaNO3 0.25 g (NH4)2SO4 0.25 g Ferric citrate monohydrate 0.20 g Na2SO4 1.00 g Na2CO3 1.25 g NaHCO3 2.50 g Na2S2O3 x 5 H2O 2.00 g Distilled water 900.00 ml Dissolve ingredients except carbonate, bicarbonate, and thiosulfate, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve carbonate and bicarbonate, adjust pH to 10 with KOH, then dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add thiosulfate to the autoclaved medium from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust the pH of the complete medium to 9-.8 10.0. MJ(-N) synthetic seawater NaCl 30.00 g K2HPO4 0.14 g CaCl2 x 2 H2O 0.14 g MgSO4 x 7 H2O 3.40 g MgCl2 x 6 H2O 4.18 g KCl 0.33 g NiCl2 x 6 H2O 0.50 mg Na2SeO3 x 5 H2O 0.50 mg Fe(NH4)2(SO4)2 x 6 H2O 0.01 g Modified W'olins mineral solution 10.00 ml Distilled water 1000.00 ml Modifi'ed Wolins mineral solution (from medium 141) Nitrilotriacetic acid 1.50 g MgSO4 x 7 H2O 3.00 g MnSO4 x H2O 0.50 g NaCl 1.00 g FeSO4 x 7 H2O 0.10 g CoSO4 x 7 H2O 0.18 g CaCl2 x 2 H2O 0.10 g ZnSO4 x 7 H2O 0.18 g CuSO4 x 5 H2O 0.01 g AlK(SO4)2 x 12 H2O 0.02 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.01 g NiCl2 x 6 H2O 0.03 g Na2SeO3 x 5 H2O 0.30 mg Na2WO4 x 2 H2O 0.40 mg Distilled water 1000.00 ml First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.
