Skip to product information
Request a Quote
Terrific Broth (TB) | High-Yield Bacterial Media | AuSaMicS
$35.00 AUD
Terrific Broth (TB)
High-yield, phosphate-buffered enriched medium for maximum biomass, plasmid DNA yield, and recombinant protein expression in E. coli and related hosts. Manufactured in Australia with full batch documentation.
AS-1429
Catalog No.
AS-1429
Form
Dehydrated powder
Final pH
7.2 ± 0.2 at 25 °C
Buffering
Potassium phosphate 0.089 M
Packaging
100 g – 5 kg
HS Code
3821.00
Overview
Terrific Broth (TB) is an enriched, phosphate-buffered bacterial growth medium specifically formulated for high-density E. coli cultivation. Developed by Tartof & Hobbs (1987), TB contains nearly double the tryptone and four times the yeast extract of standard LB Broth, combined with glycerol as a supplementary carbon source and a potassium phosphate buffer system that prevents the catastrophic pH drops that terminate growth in unbuffered media during high-density fermentation.
The result is consistently higher biomass yields, significantly greater plasmid DNA quantities per litre of culture, and more robust recombinant protein expression compared to LB or 2×YT. AuSaMicS AS-1429 is manufactured to microbiological grade standards and dispatched same-week from Melbourne with full batch documentation.
Composition (per litre of prepared medium)
| Ingredient | Concentration | Function | Mechanism |
|---|---|---|---|
| Tryptone (Casein Digest) | 12.0 g/L | Primary nitrogen & amino acid source | Pancreatic digest of casein providing free amino acids and peptides for rapid protein biosynthesis; 2× the concentration of LB Broth — sustains protein synthesis at high cell densities without nitrogen limitation |
| Yeast Extract | 24.0 g/L | Vitamins, cofactors & growth factors | Autolysate providing B-vitamins, nucleotide bases, trace minerals, and growth factors; 4× the concentration of LB — prevents vitamin limitation at OD₆₀₀ >10; critical for sustained plasmid replication and expression machinery function |
| Glycerol | 0.4% (v/v) | Non-fermentable supplementary carbon source | Slowly metabolised via gluconeogenesis without acidic fermentation by-products; prolongs exponential growth phase by supplementing glucose-free carbon; does not trigger catabolite repression — maintains cAMP-CRP regulated gene expression |
| Potassium dihydrogen phosphate (KH₂PO₄) | 0.089 M component | Buffer — pH stabilisation (acidic arm) | Conjugate acid of the phosphate buffer pair; maintains medium pH in the 7.0–7.4 range throughout high-density growth by absorbing protons generated during organic acid metabolism and glucose fermentation |
| Dipotassium hydrogen phosphate (K₂HPO₄) | 0.089 M component | Buffer — pH stabilisation (basic arm) & K⁺ source | Conjugate base of phosphate pair; donates protons to neutralise base accumulation; provides K⁺ ions for membrane potential and active transport; total phosphate 0.089 M — sufficient buffering capacity for cultures reaching OD₆₀₀ >15 |
| Final pH: 7.2 ± 0.2 at 25 °C — Slightly alkaline optimum for phosphate-buffered high-density E. coli cultivation; phosphate buffer maintains this range throughout the growth cycle | |||
Applications
Plasmid DNA productionMaximum yield of high-copy plasmid DNA per litre — miniprep, midiprep, and maxiprep workflows
Recombinant protein expressionHigh-density culture for inducible expression systems (IPTG, arabinose) — greater biomass per litre means more protein per batch
High-density fermentationShake-flask and bioreactor cultures targeting OD₆₀₀ >10 — phosphate buffer prevents pH crash at high cell densities
Competent cell preparationPre-transformation high-density culture where maximum viable cell count is needed for electroporation or chemical competency protocols
Enzyme productionHigh-yield cultivation for restriction enzymes, ligases, polymerases, and other recombinant enzyme expression systems
Antibody fragment expressionFab, scFv, and nanobody production in E. coli periplasmic and cytoplasmic expression systems
Strengths & Limitations
Strengths
Phosphate buffering — prevents pH crash in dense cultures, extending exponential growth phase significantly beyond unbuffered media
Consistently yields 2–5× more plasmid DNA per litre than LB Broth
Glycerol carbon source avoids acidic fermentation by-products that inhibit growth and protein folding
High yeast extract prevents vitamin limitation at OD₆₀₀ >10
Compatible with all standard E. coli expression strains (BL21, BL21(DE3), Rosetta, HMS174)
Full COA, TDS, SDS per batch — audit-ready documentation
Limitations
Not suitable for transformation recovery — use SOC Medium (AS-1428) post-transformation
Higher cost per litre than LB — not recommended for routine maintenance cultures
High nutrient content may support faster contaminant growth — strict aseptic technique essential
Phosphate can precipitate with divalent cations (Mg²⁺, Ca²⁺) — do not supplement with MgSO₄ or CaCl₂ directly
Not suitable for yeast or fungal cultures — use PDB (AS-1331) or MEB instead
Comparative Media — High-Density Bacterial Culture & Protein Expression
| Medium | Tryptone (g/L) | Yeast Extract (g/L) | Buffered | Carbon source | Max OD₆₀₀ | Primary use | AuSaMicS Cat. |
|---|---|---|---|---|---|---|---|
| Terrific Broth (TB) — AS-1429 | 12.0 | 24.0 | Yes (phosphate) | Glycerol 0.4% | >20 | High-density culture, plasmid & protein production | AS-1429 |
| LB Broth (Lennox) | 10.0 | 5.0 | No | None | 3–5 | Routine E. coli culture, cloning | AS-1271 |
| 2×YT Broth | 16.0 | 10.0 | No | None | 8–10 | Phage display, antibody libraries | AS-1400 |
| SOC Medium | 20.0 (casein) | 5.0 | No | Glucose (post-autoclave) | 3–5 | Post-transformation recovery only | AS-1428 |
| Auto-Induction Medium | 12.0 | 24.0 | Yes (phosphate) | Glucose + lactose | >20 | Auto-induction protein expression (no IPTG needed) | AS-1430 |
| Nutrient Broth | 5.0 (peptone) | — | No | None | 2–3 | General bacteriology, maintenance | AS-1310 |
Cross-Reference / Equivalent Products
AS-1429 is equivalent to the following products from major international suppliers:
| Supplier | Product Name | Catalogue Number |
|---|---|---|
| Oxoid (Thermo Fisher) | Terrific Broth | CM0955 |
| BD Difco | Terrific Broth | 243820 |
| Merck (Sigma-Aldrich) | Terrific Broth | T9179 |
| Thermo Fisher Scientific | Terrific Broth Modified | A1374401 |
| AuSaMicS | Terrific Broth (TB) | AS-1429 |
Preparation Instructions
1. Weigh: Suspend the required amount of AS-1429 dehydrated powder in distilled or deionised water per the label reconstitution rate.
2. Mix: Stir to dissolve completely. The broth will appear slightly turbid and golden-yellow due to the high yeast extract content.
3. Autoclave: Sterilise at 121 °C for 15 minutes (15 psi). Allow to cool before use.
4. Dispense: Distribute into sterile flasks (fill to maximum 20–25% of flask volume to ensure adequate aeration in shake-flask culture).
5. Inoculate: Inoculate with a single colony or overnight starter culture (1:50–1:100 dilution). Incubate at 37 °C with shaking at 200–250 rpm.
Storage of prepared TB: Store at 2–8 °C for up to 4 weeks. Discard if turbidity, precipitation, or colour change is observed prior to inoculation.
2. Mix: Stir to dissolve completely. The broth will appear slightly turbid and golden-yellow due to the high yeast extract content.
3. Autoclave: Sterilise at 121 °C for 15 minutes (15 psi). Allow to cool before use.
4. Dispense: Distribute into sterile flasks (fill to maximum 20–25% of flask volume to ensure adequate aeration in shake-flask culture).
5. Inoculate: Inoculate with a single colony or overnight starter culture (1:50–1:100 dilution). Incubate at 37 °C with shaking at 200–250 rpm.
Storage of prepared TB: Store at 2–8 °C for up to 4 weeks. Discard if turbidity, precipitation, or colour change is observed prior to inoculation.
💡 Pro tip — for inducible expression: Grow culture to OD₆₀₀ 0.6–0.8 before adding IPTG (typically 0.1–1.0 mM). The phosphate buffer allows continued growth post-induction, maximising protein accumulation. Lower induction temperatures (16–25 °C) improve soluble protein yield for difficult-to-fold targets.
Frequently Asked Questions
Q1: Why does Terrific Broth produce more plasmid DNA than LB?
TB contains 4× the yeast extract and 2× the tryptone of LB, providing a richer nutrient supply that supports higher cell densities (OD₆₀₀ >20 vs 3–5 in LB). More cells per litre means more plasmid DNA per preparation. The phosphate buffer also prevents the pH drop that would otherwise terminate growth mid-exponential phase in LB cultures at high density, allowing cultures to reach their maximum cell yield.
Q2: Why is glycerol used instead of glucose in Terrific Broth?
Glycerol is metabolised slowly via gluconeogenesis without generating the acidic fermentation by-products (acetate, lactate) that accumulate when glucose is present in rich media at high cell densities. These organic acids inhibit growth, destabilise plasmids, and impair protein folding. Glycerol provides a clean carbon source that extends the exponential growth phase while the phosphate buffer handles any residual pH changes.
Q3: Can Terrific Broth be used for transformation recovery?
No — TB is not recommended for post-transformation recovery. The high nutrient content and phosphate buffer are optimised for dense, growing cultures rather than the delicate recovery phase immediately after heat shock or electroporation. Use SOC Medium (AS-1428) for transformation recovery, then transfer cells to TB for expression or plasmid production cultures.
Q4: Which E. coli strains are recommended for use with Terrific Broth?
TB is compatible with all standard laboratory and expression strains including BL21(DE3), BL21(DE3)pLysS, Rosetta, HMS174(DE3), C41(DE3), C43(DE3), and DH5α. It is particularly recommended for strains used in IPTG-inducible T7 promoter expression systems where high biomass prior to induction is critical for maximal protein yield.
Q5: Can I add glucose or MgSO₄ to Terrific Broth?
Glucose addition is not recommended — it triggers catabolite repression and generates acetate at high cell densities, counteracting the benefits of the phosphate buffer. MgSO₄ and CaCl₂ should not be added directly to TB as phosphate ions precipitate with divalent cations, forming insoluble complexes that deplete the buffer capacity and reduce nutrient availability.
Important Notice: For laboratory and research use only. Not for human consumption, veterinary, food, or household use. By purchasing, the buyer confirms compliance with all applicable Australian laws, institutional biosafety requirements, and workplace health and safety regulations. AuSaMicS Pty Ltd accepts no liability for misuse outside a controlled laboratory environment.
Terrific Broth Australia | TB medium molecular biology supplier Melbourne | buy Terrific Broth E. coli high density culture | plasmid DNA production media Australia | AS-1429 AuSaMicS | high yield bacterial growth media | recombinant protein expression broth Australia | phosphate buffered bacterial media | high density E. coli culture media | Terrific Broth vs LB plasmid yield | BL21 expression media Australia | IPTG induction media high yield
Technical Data Sheet — AS-1429 Terrific Broth
Document Type
Technical Data Sheet
Product
Terrific Broth (TB)
Catalog No.
AS-1429
Revision
1.0
Issuer
AuSaMicS Pty Ltd
Technical Overview & Biochemistry
Terrific Broth was developed by Tartof & Hobbs (1987) to overcome the growth limitations of LB Broth in high-density bacterial cultivation. The two principal innovations are: (1) a greatly enriched nutrient matrix — 12 g/L tryptone and 24 g/L yeast extract — that eliminates amino acid and vitamin limitation at high cell densities; and (2) a potassium phosphate buffer system that maintains medium pH in the physiological range (7.0–7.4) throughout the growth cycle, preventing the acid accumulation that terminates exponential growth in LB at OD₆₀₀ above 3–4.
The inclusion of glycerol (0.4% v/v) as a supplementary carbon source provides a slowly metabolised, non-acidifying substrate that extends the exponential growth phase without triggering catabolite repression or generating inhibitory organic acids. Together these features enable TB cultures to routinely reach OD₆₀₀ >20 in standard shake flasks — delivering 5–10× more viable cells per litre than LB and proportionally higher yields of plasmid DNA, recombinant protein, and other biotechnological products.
Physical & Chemical Properties
| Parameter | Specification |
|---|---|
| Appearance (dehydrated powder) | Cream to golden-yellow, homogeneous, free-flowing powder |
| Appearance (prepared broth) | Golden-yellow to amber, slightly turbid solution (normal for high yeast extract content) |
| pH (prepared medium at 25 °C) | 7.2 ± 0.2 |
| Buffer system | Potassium phosphate (KH₂PO₄ / K₂HPO₄) at 0.089 M total concentration |
| Buffering capacity (pH range) | Effective pH 6.8–7.6 throughout high-density growth |
| Sterilisation | Autoclave at 121 °C, 15 min (15 psi) |
| Moisture content (powder) | ≤6% |
| Typical max OD₆₀₀ (shake flask, 37 °C) | >20 (vs 3–5 for LB) |
| HS Tariff Code | 3821.00 |
Detailed Composition (per litre)
| Ingredient | Concentration | Function | Mechanism |
|---|---|---|---|
| Tryptone (Casein Digest) | 12.0 g/L | Amino acid & nitrogen source | Pancreatic digest providing free amino acids; 2× LB concentration sustains protein synthesis at high cell densities without nitrogen limitation; rich in tryptophan (unlike soy peptone) — important for tryptophan-regulated expression systems |
| Yeast Extract | 24.0 g/L | Vitamins, cofactors & nucleotide precursors | 4× LB concentration; prevents B-vitamin depletion at OD₆₀₀ >10; provides purine/pyrimidine bases reducing nucleotide synthesis burden; trace minerals support enzyme cofactor requirements at high cell density |
| Glycerol | 0.4% (v/v) ≈ 3.2 g/L | Supplementary carbon source | Metabolised via glycerol-3-phosphate through glycolysis; does not trigger catabolite repression (no cAMP suppression); no acidic fermentation by-products; provides energy for sustained growth after tryptone carbon is exhausted |
| KH₂PO₄ | 0.089 M component ≈ 12.1 g/L | Acidic phosphate buffer arm | Accepts protons from organic acid by-products of metabolism; maintains pH above 7.0 during high-density growth; K⁺ supports membrane potential |
| K₂HPO₄ | 0.089 M component ≈ 15.5 g/L | Basic phosphate buffer arm | Donates protons when medium becomes alkaline; together with KH₂PO₄ provides ~0.089 M total phosphate — sufficient to buffer cultures to OD₆₀₀ >20 in standard shake flasks |
Mode of Action — Why TB Outperforms LB for High-Density Culture
In LB Broth, growth terminates at OD₆₀₀ 3–5 due to two simultaneous limiting factors: nutrient exhaustion (amino acids and vitamins depleted) and pH drop below 6.5 from accumulated organic acids (acetate, formate). TB addresses both: the enriched nutrient matrix (4× yeast extract, 2× tryptone) delays nutrient limitation to OD₆₀₀ >15, while the 0.089 M potassium phosphate buffer absorbs protons from organic acid metabolism, maintaining pH 7.0–7.4. Glycerol provides additional carbon without acidification. The combined effect extends the exponential growth phase by 6–10 hours in standard shake-flask culture, resulting in 5–10× higher final cell density and proportionally higher yields of all intracellular products.
Quality Control — Performance Test Organisms
| Organism | ATCC / Reference | Inoculum (CFU) | Incubation | Expected Result | Status |
|---|---|---|---|---|---|
| Escherichia coli K-12 | ATCC 10798 | ≤100 CFU | 37 °C, 200 rpm, 18 h | Good growth; OD₆₀₀ ≥ reference TB lot | PASS |
| Escherichia coli BL21(DE3) | ATCC BAA-1025 | ≤100 CFU | 37 °C, 200 rpm, 18 h | OD₆₀₀ >15 (shake flask, 250 mL / 50 mL fill) | PASS |
| Sterility check (uninoculated) | — | — | 37 °C, 48 h | No growth / no turbidity increase | PASS |
Literature & References
- Tartof, K.D. & Hobbs, C.A. (1987). Improved media for growing plasmid and cosmid clones. Bethesda Research Laboratories Focus, 9, 12.
- Sambrook, J. & Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press. Appendix 1.
- Green, M.R. & Sambrook, J. (2012). Molecular Cloning: A Laboratory Manual, 4th edn. Cold Spring Harbor Laboratory Press.
- Studier, F.W. (2005). Protein production by auto-induction in high-density shaking cultures. Protein Expression and Purification, 41(1), 207–234.
- Rosano, G.L. & Ceccarelli, E.A. (2014). Recombinant protein expression in Escherichia coli: advances and challenges. Frontiers in Microbiology, 5, 172.
Download TDS Document
AuSaMicS Pty Ltd Disclaimer: Technical data is based on current literature and internal quality testing. Values may vary by lot. For laboratory use only.
Safety Data Sheet — AS-1429 Terrific Broth
Full 16-section Safety Data Sheet compliant with GHS and Australian Work Health and Safety (WHS) Regulations. Download the complete SDS before handling this product.
GHS Standard
GHS Rev. 9
WHS Compliance
Australian WHS Reg. 2017
Sections
16 mandatory
Signal Word
Not classified
SDS Section Summary
| Section | Heading | Key Information |
|---|---|---|
| 1 | Identification | Terrific Broth, AS-1429, AuSaMicS Pty Ltd, support@ausamics.com, +61 412 520 598 |
| 2 | Hazard identification | Not classified as hazardous under GHS. No signal word. Dust may cause mild respiratory irritation. |
| 3 | Composition / ingredients | Tryptone, Yeast Extract, Glycerol, KH₂PO₄, K₂HPO₄ — all non-hazardous biological/food-grade materials |
| 4 | First aid | Inhalation: fresh air. Skin/eyes: wash with water 15 min. Ingestion: rinse mouth; seek advice if unwell. |
| 5 | Firefighting | Not flammable. CO₂ or dry powder extinguisher. SCBA in enclosed fire scenarios. |
| 6 | Accidental release | Sweep up powder; avoid dust generation. Prepared broth: absorb with inert material; dispose per local regulations. |
| 7 | Handling & storage | Store powder at 2–25 °C, sealed, dry. Prepared medium at 2–8 °C, use within 4 weeks. |
| 8 | Exposure controls / PPE | Lab coat, nitrile gloves, safety glasses. No respiratory protection required for normal use. |
| 9 | Physical & chemical properties | Golden-yellow powder (dry); amber liquid (prepared). Not flammable. pH 7.2 prepared. |
| 10 | Stability & reactivity | Stable. Incompatible with strong oxidisers. No hazardous decomposition under normal conditions. |
| 11 | Toxicological information | Not acutely toxic. All components have low oral, dermal, and inhalation toxicity profiles. |
| 12 | Ecological information | Biodegradable. Not environmentally hazardous. Avoid large releases to waterways (high BOD). |
| 13 | Disposal | Autoclave inoculated broth before disposal. Uninoculated: dilute and drain. Dry powder: general lab waste. |
| 14 | Transport | Not classified as dangerous goods (UN/ADG). No special transport requirements. |
| 15 | Regulatory | Not subject to AICIS notification. Complies with Australian WHS model regulations. |
| 16 | Other information | Issued by AuSaMicS Pty Ltd. For laboratory use only. |
Download Full SDS (16-Section, GHS-Compliant)
AuSaMicS Pty Ltd Safety Disclaimer: Always consult the full SDS before handling. Use appropriate PPE. Emergency contact: +61 412 520 598.
Certificate of Analysis — AS-1429 Terrific Broth
A batch-specific Certificate of Analysis is issued for every production lot of AS-1429. The COA confirms that the supplied lot meets all specifications below. Retain for laboratory audit and accreditation purposes.
Issued by
AuSaMicS Pty Ltd
Test frequency
Every production lot
Traceability
LOT & EXP on label
Dispatch
Included with order
Specification Table
| Test Parameter | Method | Specification | Typical Result | Status |
|---|---|---|---|---|
| Appearance (powder) | Visual | Cream to golden-yellow, homogeneous, free-flowing | Golden-yellow, homogeneous | PASS |
| Appearance (prepared broth) | Visual | Golden-yellow to amber, slightly turbid | Amber, slightly turbid | PASS |
| pH (prepared, 25 °C) | Potentiometry | 7.2 ± 0.2 | 7.2 | PASS |
| Moisture content | Loss on drying | ≤6.0% | 4.1% | PASS |
| Growth promotion — E. coli K-12 ATCC 10798 | OD₆₀₀ / incubation | OD₆₀₀ ≥ reference TB lot at 18 h | OD₆₀₀ >15 at 18 h, 37 °C, 200 rpm | PASS |
| pH stability post-growth (high-density) | Potentiometry post-culture | pH ≥ 6.8 at OD₆₀₀ = 15 | pH 7.0 at OD₆₀₀ 15+ | PASS |
| Sterility (prepared medium) | Incubation 37 °C / 48 h | No growth | No growth | PASS |
Download Sample COA
AuSaMicS Pty Ltd COA Disclaimer: This Certificate of Analysis is lot-specific. The sample COA is representative only. The actual COA for the supplied lot is included with every order. For lot-specific documentation: support@ausamics.com.