# 1560: MARINES PLANCTOMYCES MEDIUM ## Main sol. 1560 0.25 g Peptone (Bacto) 0.25 g Yeast extract (Bacto) 2.38 g HEPES 20 ml **Mineral salts solution** 1 ml **Trace element solution** 250 ml **Artifical sea water** 15 g Agar (Bacto) 730 ml Double distilled water 1. Sterilize 20 min./121°C. After cooling to 60°C add: 8 ml N-Acetylglucosamine (10% stock solution, 10 % stock solution) 10 ml Glucose (2.5%, sterile-filtered) 5 ml **Vitamin solution (double conc.)** (medium 621) 2. Adjust pH to 7.5 (the medium is only weakly buffered; one needs approx. 10 drops/l medium of 6 N KOH). ## Artifical sea water 46.94 g NaCl 7.84 g Na2SO4 21.28 g MgCl2 x 6 H2O 2.86 g CaCl2 0.38 g NaHCO3 1.38 g KCl 0.192 g KBr 0.052 g H3BO3 0.08 g SrCl2 6 mg NaF 1000 ml Double distilled water ## Mineral salts solution 10 g Nitrilotriacetic acid (NTA) 29.7 g MgSO4 x 7 H2O 3.34 g CaCl2 x 2 H2O 12.67 mg Na2MoO7O4 x 2 H2O 99 mg FeSO4 x 7 H2O 50 ml **"Metals 44"** 950 ml Distilled water 1. Dissolve the nitrilotriacetic acid, adjust the pH to 7.0 with KOH (about 7.3 g). Dissolve other salts separately, combine and adjust the pH to 6.8 with NaOH or H2SO4. ## "Metals 44" 250 mg Na-EDTA 1095 mg ZnSO4 x 7 H2O 500 mg FeSO4 x 7 H2O 154 mg MnSO4 x H2O 39.2 mg CuSO4 x 5 H2O 24.8 mg Co(NO3)2 x 6 H2O 17.7 mg Na2B4O7 x 10 H2O 1000 ml Distilled water ## Trace element solution 1.5 g Na-Nitrilotriacetat 0.5 g MnSO4 x H2O 0.1 g Fe SO4 x 7 H2O 0.1 g Co(NO3)2 x 6 H2O 0.1 g ZnCl2 0.05 g NiCl2 x 6 H2O 0.05 g H2SeO3 0.01 g CuSO4 x 5 H2O 0.01 g AlK(SO4)2 x 12 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.01 g Na2WO4 x 2 H2O 1000 ml Double distilled water ## Vitamin solution (double conc.) 4 mg Biotin 4 mg Folic acid 20 mg Pyridoxine hydrochloride 10 mg Riboflavine 10 mg Thiamine-HCl x 2 H2O 10 mg Nicotinamide 10 mg D-Calcium pantothenate 0.2 mg Vitamin B12 10 mg p-Aminobenzoic acid 1000 ml Double distilled water