Terrific Broth (1 L)
Terrific broth is a rich medium for E. coli that incorporates glycerol as an energy source, resulting in accelerated bacterial growth and increased yield per volume. Moreover, it contains potassium phosphates to mitigate the risk of cell death caused by pH fluctuations. Terrific broth is used to propagate E. coli prior to infection by filamentous phage.
Reagents
12 g Bacto-tryptone
24 g Yeast extract
4 mL (5.04 g) Glycerol
2.31 g KH2PO4
12.54 g K2HPO4
Protocol
Potassium Phosphate Buffer
- Dissolve 2.31 g KH2PO4 and 12.54 g K2HPO4 in 90 mL water.
- Adjust the volume to 100 mL with water and autoclave. Store at room temperature.
Terrific Broth
- Dissolve 12 g bacto-tryptone, 24 g yeast extract, and 4 mL (5.04 g) glycerol in 900 mL water.
- Divide into 10 bottles with 90 mL in each and autoclave.
- When cooled, add to each bottle add 10 mL of the separately autoclaved potassium phosphate buffer.
- Store at room temperature.