L-Broth (Luria Broth) is the standard nutrient media for propagation of Escherichia coli for purposes of strain maintenance, cloning, plasmid propagation, and protein expression. LB Broth media formulations have been industry standards for the cultivation of Escherichia coli since the 1950’s (1,2,3). These media have been widely used in molecular microbiology applications for the preparation of plasmid DNA and recombinant proteins (4,5). The media are nutrient-rich formulations which provide peptides and peptones, vitamins, and trace elements. The three formulations (Lennox, Miller and Luria) differ in the amount of sodium chloride, thus providing selection of the appropriate osmotic conditions for the particular bacterial strain and desired culture conditions. The low salt formulations, Lennox and Luria, are ideal for cultures requiring salt-sensitive antibiotics such as Zeocin™.
Dissolve 15.5 grams of the powder in one liter of ddH2O, heating with stirring until solubilized. Dispense into containers, loosen caps and autoclave for 15 minutes at 121ºC (15psi).
Store powdered media at RT. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept refrigerated and used within a short period of time.
| Component | g/liter |
|---|---|
| Tryptone | 10 |
| Yeast Extract | 5 |
| Sodium Chloride | 0.5 |
| Total: 15.5g/Liter | |
| 500g makes 32.25 liters |
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