Tartoff - Hobbs Broth (Terrific Broth)

Tartoff-Hobbs (Terrific) Broth is recommended for the cultivation of recombinant strains of Escherichia coli.

Composition**

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 47.6 grams in 1000 ml distilled water containing 4 ml glycerol. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Mix well and dispense as desired.

Principle And Interpretation

Tartoff Hobbs developed the Terrific Broth Medium for cultivation of recombinant Escherichia coli strains. These strains have extended growth phase when cultivated in this medium (1). Tartoff-Hobbs Medium supports high cellular density and mass and maintains the growth in the logarithmic phase for a long time. Due to this fact, it provides greater yields of recombinant proteins and plasmid DNA. Often, Tartoff-Hobbs (Terrific) Broth substitutes Luria Bertani Broth (M1151), to get enhanced yields of plasmid DNA and recombinant proteins. The procedures for inoculation, incubation and generation of recombinant strains are detailed by Sambrook et al (2).

Casein enzymic hydrolysate and yeast extract supply the necessary nutrients and cofactors for the excellent growth of recombinant strains of E. coli. The addition of extra peptone and yeast extract in the medium allows higher plasmid yield per volume. The two phosphates provide good buffering action to the medium. Glycerol is used as the carbohydrate source. Unlike glucose, glycerol is not fermented to acetic acid.

Quality Control

Appearance Cream to light yellow homogeneous free flowing powder

Colour and Clarity of prepared medium Light amber coloured clear solution without any precipitate.

Reaction Reaction of 4.76% w/v aqueous solution at 25°C. pH: 7.2±0.2

pH 7.00-7.40

Cultural Response

M1250: Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours. (Growth evident by turbidity)

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label.

Reference

1. Tartoff K. D. and Hobbs C. A., 1987, Improved Media for Growing Plasmid and Cosmid Clones, Bethesda Res. Lab. Focus, 9:12.
2. Sambrook J., Fritsch, E. E., and Maniatis T., 1989, Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Lab., Cold Spring Harbor, N.Y.