f/2 vitamin solution

Reference: Guillard, R.R.L. and J.H. Ryther.  1962.  Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea Cleve. Can. J. Microbiol. 8: 229-239.

Reference: Guillard, R.R.L.  1975.  Culture of phytoplankton for feeding marine invertebrates in “Culture of Marine Invertebrate Animals.”  (eds: Smith W.L. and Chanley M.H.)  Plenum Press, New York, USA.  pp 26-60.

Substock solutions:

First prepare separate substock solutions of cyanocobalamin (Vitamin B12) and biotin in purified water (e.g., Milli-Q, Nanopure, distilled water).

Substock Substance Concentration
1. Vitamin B12 (Cyanocobalamin) 10 mg/10 mL
2. Biotin 5 mg/50 mL

Working stock solution:

Then prepare the final working solution by combining the following amounts of the two substock solutions plus thiamine HCl to 100 mL of purified water.

Substance Concentration (per 100 mL)
1.  Vitamin B12 (Cyanocobalamin) 0.1 mL
2.  Biotin 1.0 mL
3.  Thiamine HCl 20 mg

Dispense the working solution as 1 mL aliquots in cryovials for convenient use when preparing the medium. Store the remainder of the primary stocks and working solution in polyethylene bottles. Wrap vials/bottles with Parafilm to avoid moisture loss and store all solutions frozen.