The macronutrients are removed from the stock bottles and combined in a clean beaker. Micronutrients are all found in one stock bottle and a total amount relative to the amount of media desired is obtained. Iron, vitamins, organics, and distilled water are then added to the mixture.
Appropriate carbon is added and mixed by method of a stir-bar.
While the solution is mixing, the pH is adjusted to 5.7 using a pH meter and an acid and a base as appropriate. The magnetic stir-bar is removed and the agar is added. The media is then microwaved until the agar is dissolved, but not to boiling. The clear, liquid media is distributed evenly across test tubes and the caps are placed on the tubes and labeled. The media is sterilized in an autoclave for fifteen minutes at 15lbs pressure.