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.