Terrific Broth, TB media, TB Medium w/o Glycerol | 250g
AUD 75.00
Excl.GST
- Product Code: GMNB-TBM116
- Availability: In Stock
Tags: TB Medium
TB culture media
Recipe same as the Cold spring harbour protocol*, with more nitrogen source and buffer capability.
250g, fine powder, up to make 4.25L liquid media.
500g, fine powder, up to make 8.5L liquid media.
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
KH2PO4 0.017 M
K2HPO4 0.072 M
Preparation
- Suspend 58.8 g of the medium into distilled or MQ water.
- [optional] Add 4 ml Glycerol (prepared separately by customer,up to 8 ml if prolonged culture is needed).
- Mix well with continuous stirring and make up to 1000 ml.
- Sterilise by autoclaving at 121°C for 15 minutes. Aliquot as needed.
Caution
The TB powder may form small crystals when exposed to air due to moisture absorption of K2HPO4. These crystals can be weighed along with the powder and dissolved in water without affecting the preparation of the media.
*Ingredients difference between protocols
| Ingredients | Tryptone | Yeast extract | K2HPO4 | KH2PO4 | Glycerol |
|---|---|---|---|---|---|
| CSHL | 20 gm | 24 gm | 12.5 gm | 2.3 gm | 4 ml |
| MERCK / Thermo Fisher | 11.8 gm | 23.6 gm | 9.4 gm | 2.2 gm | 4 ml |
