Terrific Broth, Liquid TB Medium, with Glycerol |500ml

Liquid TB media,  Recipe is the same as the Cold spring harbour protocol*, with more nitrogen source and buffer capability.

500ml liquid media.

Introduction

Terrific broth is a buffered and enriched medium used to cultivate recombinant strains of E.coli. It is a nutrient-rich medium containing glycerol, peptides, amino acids, water-soluble vitamins, and carbohydrates. Terrific broth (TB) contains more tryptone and yeast extract than the standard LB medium. Terrific broth can be used for almost all applications involving liquid bacterial culturing steps such as protein expression and cloning.

Principle

The terrific broth is a nutrient-rich growth medium that supports the high cell density of E.coli. It maintains growth in the logarithmic phase resulting in larger yields of plasmid DNA and heterologous proteins. Tryptone and yeast extract act as the sources of carbon, nitrogen, vitamins, and amino acids required for the growth of bacteria. The potassium phosphate buffer system acts as the source of potassium and prevents cell death and, eventually, the drop in pH. Glycerol is added to the medium instead of glucose as a carbon source. Glycerol is not reduced to acetic acid. Glucose acts as a repressor in various protein expression systems, whereas glycerol promotes growth.

Applications

It gives a higher plasmid yield per volume compared with LB media.

TB (not to be confused with T broth) can obtain higher yields of recombinant protein expression in E.coli. However, some proteins require slow E.coli growth for proper folding. For such cases, it would be better to use M9 media.

Components

Terrific Broth (TB) medium, per litre

Yeast extract (premium)      24 g

Tryptone              20 g

Glycerol               4 mL

KH2PO4                0.017 M 

K2HPO4                0.072 M

Preparation

Use directly after opening, the remaining liquid should be sealed immediately and stored in the 4°C refrigerator. Operation on a sterile bench is recommended.

*Ingredients difference between protocols