Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use:
Recommended for the cultivation of recombinant strains of Escherichia coli.
Composition**
| Ingredients | Gms/Litre |
|---|---|
| Tryptone | 12.000 |
| Yeast extract | 24.000 |
| Potassium dihydrogen phosphate | 2.200 |
| Dipotassium hydrogen phosphate | 9.400 |
| Final pH (at 25°C) | 7.2±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Each capsule contains 11.9 gms of medium. Suspend 1 capsule in 250 ml (4 capsules in 1000 ml) distilled or purified water containing 0.4% v/v glycerol (i.e. 1 ml in 250ml or 4 ml in 1000ml media). Heat to boiling to dissolve the medium completely. Mix well and dispense into tubes or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
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 (4). 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 HiEncap™ Luria Bertani Agar, Miller (EC1151CCL/EC1151D), 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 (3).
Tryptone 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.
Type of specimen
Isolated Microorganism from recombinant strains samples.
Specimen Collection and Handling:
For isolated Microorganism samples follow appropriate techniques for handling specimens as per established guidelines (4). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations :
- This medium is general purpose medium and may not support the growth of fastidious organisms.
- Further biochemical and serological tests must be carried out for further identification.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance: Gelatin capsule containing cream to light yellow coloured granular media
Colour and Clarity of prepared medium: Light amber coloured clear solution without any precipitate.
Quantity: Each capsule contains 11.9 grams medium sufficient for 250 ml media.
Reaction: Reaction of 4.76% w/v aqueous solution at 25°C. pH: 7.2±0.2
pH: 7.00-7.40
Cultural Response: Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours. (Growth evident by turbidity)
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Escherichia coli ATCC 23724 | 50-100 | good |
| Escherichia coli ATCC 39403 | 50-100 | good |
| Escherichia coli ATCC 47014 | 50-100 | good |
| Escherichia coli ATCC 53868 | 50-100 | good |
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (1,2).
| Product Name | HiEncap™ Terrific Broth (HiEncap™ Tartoff -Hobbs Broth) |
|---|---|
| SKU | EC1250CCL |
| Product Type | HiEncap™ |
| Physical Form | Granulated Media in Gelatin Capsule |
| Origin | Animal |
| Packaging type | HDPE Jar |
| References | 1. Tartoff K. D. and Hobbs C. A., 1987, Improved Media for Growing Plasmid and Cosmid Clones, Bethesda Res. Lab. Focus, 9:1 2. 2. Sambrook J., Fritsch, E. E., and Maniatis T., 1989, Molecular Cloning : A Laboratory Manual, 2nd Ed., Cold Spring HarborLab., Cold Spring Harbor, N.Y. |
| Customized Product Available | No |
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