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Spirulina-Medium

Lit.: modified from Aiba, S. & Ogawa, T. (1977): Assessment of growth yield of a blue-green alga: Spirulina platensis, in axenic and continuous culture. J. Gen. Microbiol. 1977, 102: 179-182.

For 1000 mL final culture medium: Stock Solution SL SL I Volume 500 mL Component NaHCO3 Na2CO3 K2HPO4 NaNO3 K2SO4 NaCl MgSO4 . 7H2O CaCl2 . 2H2O FeSO4 . 7H2O EDTA (Titriplex III) micronutient solution* Concentration SL 13.61 g 500 mL-1 4.03 g 500 mL-1 0.50 g 500 mL-1 2.50 g 500 mL-1 1.00 g 500 mL-1 1.00 g 500 mL-1 0.20 g 500 mL-1 0.04 g 500 mL-1 0.01 g 500 mL-1 0.08 g 500 mL-1 5.0 mL 500 mL-1 Conc. Final Medium 4.05 10-2 M 9.50 10-3 M 7.17 10-4 M 7.35 10-3 M 1.43 10-3 M 4.27 10-3 M 4.15 10-4 M 9.01 10-5 M 1.64 10-5 M

results in a pH of 9.09.2 SL II 500 mL

results in a pH of pH = 3.84.0 Autoclave solutions I and II separately, unite after cooling and add 5 10-6 g L-1 vitamin B12 in sterile solution, if required (usually vitamins are only required for long-term cultures/stock cultures, however, some algal species require some vitamins). Stock solutions should be stored at 4 C. Final pH after combining SL I + SL II = 9.29.5. For agar slants: 1. Prepare solution I and II as double strength solution, i.e. above amounts of salts in 250 mL each. Autoclave each solution separately. 2. Prepare agar solution in double strength (for 1 L final solution, 1 % agar): - 500 mL distilled water + 10 g Agar Agar (use 1 L bottle at minimum) - heat in microwave to dissolve - autoclave together with solutions I + II - let cool to +50+60 C 3. Combine 250 mL each of autoclaved solutions I and II and autocalved agar solution - mix and add vitamin B12 aseptically When preparing liquid medium from double strength stock solutions I and II, autoclave distilled water separately and, after cooling, combine 1 part stock I + 1 part stock II + 2 parts distilled water, i.e. 250 mL + 250 mL + 500 mL. Finally add vitamin B12 from stock solution and dispense in sterile culture tubes, flasks - let stand for a minimum of 3...5 days and inoculate from algal stock cultures. * Preparation of the micronutrient solution Solution I 881mL 1 mL 2 mL 5 mL 5 mL 5 mL 1 mL 0.4 g Soution II 100 mL 0.7 g 0.4 g Solution I and II - autoclave separately - let cool down - pool together

CCCryo 4/2010

distilled water ZnSO4 . 7H2O MnSO4 . 4H2O H3BO3 Co(NO3)2 . 6H2O Na2 MoO4 .2H2O CuSO4 . 5H2O EDTA

distilled Water FeSO4 7H2O EDTA