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Originally from Tartoff, et al. (1987), printed in Cold Spring Harbor Protocol.
Characterized with LB in Danquah & Forde (2007).  

Plasmid yield is ≈10 µg/mL for pUC19, fourfold more than LB, but at 65% the medium cost per amount plasmid, according to Danquah & Forde.
Shyam finds that E. coli TG1 saturates at OD600 ≈ 18–20 in and at OD600 ≈ 6 in LB.

Terrific Broth

  • 12 g/L tryptone (1.2% m/V)
  • 24 g/L yeast extract (2.4% m/V)
  • 4 mL/L glycerol (4.34 mM, 0.4% V/V)
  • 17 mM KH2PO4, 2.31 g/L
  • 72 mM K2HPO4, 12.54 g/L
  • (opt.) 15 g/L agar (1.5% m/V)
  • (opt.) 5–10 mM 1 M MgSO4 or MgCl2 (nontraditional)

It was originally suggested to separately autoclave phosphate buffer and agar. It was recently found that phosphate and agar react to form H2O2, but E. coli are less, if at all, significantly impacted due to catalase expression. Commercial mixes do not separate the two, yet instruct autoclaving. There is also reason for separately autoclaving the glycerol, as glycerol partakes in Maillard reactions with tryptone, though probably less so than sugars do. Maillard reactions sequester nutrients and form products that decompose into growth-inhibitory compounds.

Magnesium is deficient in LB, but high level of trace mineral, including Mg2+, contamination of phosphate salts might provide enough in TB. To be sure, 10 mM MgSO4 or MgCl2 can be added, but is not part of the traditional TB recipe.

Recovery Medium:

Shyam rationalizes that TB supplemented with 0.4% V/V glucose and MgCl2 and/or MgSO4 to a final 10–20 mM would be just as good as SOC for transformation recovery. SOB/SOC, has 10 mM MgCl2 and 10 mM MgSO4 to help rebuild the cation-associated E. coli LPS.

Procedure for 1 L

  1. Dissolve and autoclave on liquid cycle, 20 min, 15 psi:
    1. 12 g tryptone
    2. 24 g yeast extract
    3. 4 mL glycerol (or 8 mL 50% glycerol)
      (or add after autoclaving) 
    4. (opt.) 15 g agar
    5. (opt.) 5–10 mL 1 M MgSO4 or MgCl2
    6. final 900 mL diH2O
  2. Prepare or obtain 100 mL 10× TB buffer (0.17 M KH2PO4 0.72 M K2HPO4)
    Dissolve and autoclave on liquid cycle, 20 min, 15 psi:
    1. 2.31 g KH2PO4
    2. 12.54 g K2HPO4
    3. ≈90 mL diH2O, adjusted to 100 mL final
  3. When 900 mL nutrient solution has cooled ≤60°C, add 100 mL sterile buffer.
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