Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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Hofer’s Alkaline Medium
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$57.20
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SKU:
M717
Used for selective isolation of Agrobacterium species while inhibiting Rhizobium species from soil samples.
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Intended Use
Recommended for selective isolation of Agrobacterium species while inhibiting Rhizobium species from soil samples.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Mannitol | 10.000 |
| Dipotassium hydrogen phosphate | 0.500 |
| Magnesium sulphate | 0.200 |
| Sodium chloride | 0.100 |
| Yeast extract | 1.000 |
| Thymol blue | 0.016 |
| Agar | 15.000 |
| Final pH (at 25°C) | 11.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 26.8 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Agrobacterium is a genus of bacteria that causes tumours in plants. Most strains of Agrobacterium are plant pathogens and their natural habitat is on and around the roots and underground stems of susceptible plants (1). Agrobacterium tumefaciens is the most commonly studied species in this genus. Agrobacterium is well known for its ability to transfer DNA between itself and plants, and for this reason it has become an important tool for plant improvement by genetic engineering. Hofers Alkaline Medium is formulated as described by Subba Rao (4) and is used for growing Agrobacterium species while inhibiting Rhizobium species from soil. It is a selective medium with high alkaline pH. Agrobacteria grow at higher pH while Rhizobia fail to grow at alkaline pH. The medium is supplemented with mannitol as the carbohydrate or carbon source. Yeast extract provides nitrogenous nutrients.
Sodium chloride maintains osmotic balance of the medium. Dipotassium phosphate buffers the medium. Thymol blue is the pH indicator, which remains blue at high alkaline pH.
Type of specimen
Soil samples
Specimen Collection and Handling
For soil samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(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
- 1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- 2. 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
Light yellow to light green homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Blue coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 2.68% w/v aqueous solution at 25°C. pH : 11.0±0.2
pH
10.80-11.20
Cultural Response
Cultural characteristics observed after an incubation at 25-28°C for upto 5 days.
| Organism | Growth |
|---|---|
| Agrobacterium luteum ATCC 25657 | good-luxuriant |
| Agrobacterium tumefaciens ATCC 15955 | good-luxuriant |
| Rhizobium trifolii ATCC 14480 | inhibited |
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (2,3).
| Product Name | Hofer’s Alkaline Medium |
|---|---|
| SKU | M717 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal Free (Microbial) |
| Packaging type | HDPE |
| References | 1.Balows A., Truper H. G., Dworkin M., Harder W., Scheifer K. H., (Eds.), The Prokayotes, 2nd Edition, Springer- Verlag,New York Inc. 2.Subba Rao N. S., 1977, Soil Microorganisms and Plant Growth, Oxfordand IBH Publishing Co., New Delhi. |
| Customized Product Available | No |
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