Terrific Broth (TB) Medium Preparation and Recipe

To prepare L  Terrific Broth (TB) Medium

Table 1. Recipe

1. Prepare 0.8 L of distilled water in a suitable container.
2. Add 12 g of tryptone to the solution.
3. Add 24 g of yeast extract to the solution.
4. Add 4 mL of glycerol to the solution.
5. Add 2.313 g of Potassium dihydrogen phosphate to the solution.
6. Add 12.54 g of Potassium phosphate dibasic to the solution.
7. Add distilled water until the volume is 1 L.
8. The original suggestion was to autoclave phosphate buffer and agar separately. However, it has recently been discovered that when combined, phosphate and agar react to produce H2O2. Fortunately, this reaction does not greatly affect E. coli due to their high catalase expression. Despite this, commercial mixes do not separate the two components and still recommend autoclaving. It is also worth noting that autoclaving glycerol separately is necessary because it can participate in Maillard reactions with amines found in tryptone/yeast extract. Although glycerol has fewer carbonyl groups, which contribute to Maillard reactions, it can still sequester nutrients and form growth-inhibitory compounds.