TERRIFIC BROTH

Features

Quantity: 500g
Appearance: Beige powder. Autoclaved medium should be amber.
Storage: 2ºC – 25ºC. When not in use, keep container closed to avoid hydration.

Formulation (g/L)
Tryptone: 12,00
Yeast Extract: 24,00
K2HPO4: 12,54
KH2PO4: 2,31
Final pH (25ºC): 7,2 ± 0,2

Preparation:
Add 47,6g of the dehydrated medium to 900 ml of distilled water. Mix well and add 4 ml of glycerol. Adjust final volume to 1 liter. Dissolve by heating with regular agitation. Boil for 1 minute in order to dissolve  completely. Dispense in appropriate containers and sterilize by autoclaving at 121ºC for 15 to 20 minutes. Store at 2ºC to 8ºC.

Supplements
Terrific Broth is a rich growth medium, which contains all the nutritional requirements for E.coli to support a high cell density and maintaining growth in the logarithmic phase for an extended period of time, resulting in larger yields of plasmid DNA and heterologous proteins. Tryptone and Yeast Extractare the sources for carbon, nitrogen, vitamins, minerals, and amino acids essential for growth,  whereas the potassium phosphate buffer system acts to prevent cell death. Glycerol serves an additional source for carbon as well as a source for carbohydrates with the advantage over glucose that it is not fermented into acetic acid. Moreover, in many protein expression experiments, glucose acts as a repressor. Other supplements, including antibiotics, can be added. As many are heatsensitive, they cannot be autoclaved and should be filter-sterilized prior to adding to the medium it has cooled down.