SN Medium (Waterbury 1986)
Filter 750 mL seawater through a 0.22 uM filter and add 250 mL distilled H2O. Autoclave in a teflon lined bottle then add the following compounds aseptically:
Quantity Compound Stock Solution 2.5 mL NaNO3 300.0 g/L dH2O 2.6 mL K2HPO4 (anhydrous) 6.1 g/L dH2O 5.6 mL Na2EDTA.2H2O 1.0 g/L dH2O 2.6 mL Na2CO3 4.0 g/L dH2O 1.0 mL Vitamin B12 (Cyanocobalamin) 1.0 mg/L dH2O 1.0 mL Cyano Trace Metal Solution (see recipe below)
Modified SN Medium (CCMP Recipe)
CCMP modifies Waterbury's SN Medium as follows: 1.0 L filtered seawater is autoclaved in a in teflon lined bottle and the following is added aseptically:
Quantity Compound Stock Solution 10.0 mL NaNO3 300.0 g/L dH2O 2.5 mL K2HPO4 (anhydrous) 6.1 g/L dH2O 5.0 mL Na2EDTA.2H2O 1.0 g/L dH2O 2.5 mL Na2CO3 4.0 g/L dH2O 1.0 mL H2SeO3 1.29 mg/L dH2O 1.0 mL Cyano Trace Metal Solution (see recipe above) 1.0 mL Va Vitamin Solution (see recipe above)
CYANO TRACE METAL SOLUTION
The original SN trace metals are made by dissolving each metal compound individually in 100 mL dH2O. All six solutions are then combined by bringing the volume up to 1.0L with dH2O. Filter sterilize the solution and store in 50cc Corning tubes at -20deg C.
Quantity Compound 6.25 g Citric Acid*H2O 6.0 g Ferric Ammonium Citrate 1.4 g MnCl2*4H2O 0.39 g Na2MoO4*2H2O 0.025 g Co(NO3)2*6H2O 0.222 g ZnSO4*7H2O
CYANO TRACE METAL SOLUTION WITHOUT NITROGEN
For experiments on nitrogen metabolism I frequently replace the nitrogen containing trace metal salts in the SN cyano trace metal solution with non-nitrogenous equivalents. I call these 'SN Cyano Trace-N' and prepare them as above with the following ingredients:
Quantity Compound 6.25 g Citric Acid*H2O 4.26 g Ferric Chlroide (FeCl3*6H2O) 1.4 g MnCl2*4H2O 0.39 g Na2MoO4*2H2O 0.022 g CoCl2*6H2O 0.222 g ZnSO4.7H2O
VA Vitamin mix (Davis and Guillard, 1958)
To 60 mL of dH2O add:
Quantity Compound Stock Solution 100 mg Inositol - 20 mg Thiamine • HCl - 0.1 mL Vitamin B12 (cyanocobalamin) 1 g/L dH2O 1 mL Biotin 0.1 g/L dH2O 0.1 mL Folic acid 2 g/L dH2O 0.5 mL p-aminobenzoic acid 2 g/L dH2O 10 mL Niacin 1 g/L dH2O 10 mL Ca-pantothenate 2 g/L dH2O 10 mL Pyridoxine 1 g/L dH2O
Bring volume up to 100 ml and filter sterilize. Store in 50cc Corning tubes in freezer.
The original references for this medium are:
Waterbury, J.B., Watson, S.W., Valois, F.W. and Franks, D.G. 1986. Biological and ecological characterization of the marine unicellular cyanobacterium Synechococcus. pp 71-120. In (Platt, T. and Li, W.K.W. eds.) Photosynthetic Picoplankton. Can. Bull. Fish. Aquatic Sci. 214. Department of Fisheries and Oceans, Ottawa, Canada.
Davis, H.C. and Guillard, R.R.L 1958. Relative value of ten genera of micro-organisms as foods for oyster and clam larvae. U.S.F.W.S. Fish Bull. 136 (58): 293-304.