HiEncap™ Luria Agar Base, Miller's Modification

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EC1726CCL
For the cultivation and maintenance of recombinant strains of Escherichia coli with or without addition of glucose.


Intended Use:

Recommended for the cultivation and maintenance of recombinant strains of Escherichia coli with or without addition of glucose.

Composition**

Ingredients Gms / Litre
Tryptone 10.000
Yeast extract 5.000
Sodium chloride 0.500
Agar 15.000
Final pH (at 25°C) 7.0±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Each capsule contains 7.63 gms of medium. Suspend 1 capsule in 250 ml (4 capsules in 1000 ml) distilled or purified water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. If desired add filter sterilized 20% v/v glucose solution i.e 2.5 ml in 250 ml or 10 ml in 1000 ml media. Mix well and pour into sterile Petri plates.

Principle And Interpretation

This medium is based on original formula described by Miller for the growth and maintenance of E.coli strains used in molecular microbiology (4). Luria Agar Base, Miller is a nutritionally rich medium recommended for growth of pure cultures of recombinant strains. E.coli is grown in late log phase in LB medium. Some plasmid vectors may replicate to high copy numbers without selective amplification. Some vectors do not replicate so freely, and need to be selectively amplified. Chloramphenicol can be added to inhibit host synthesis and as a result prevent replication of the bacterial chromosome. (5) Luria Agar Base, Miller's modification contains one tenth and one twentieth the sodium chloride level of the Lennox and Miller formulations of LB Agar respectively (3,4,5). This helps the user to select the optimal salt concentration for a specific strain. The medium may be aseptically supplemented with glucose, if desired. Tryptone provides peptides and peptones while Vitamin B complex is provided by yeast extract. Sodium chloride provides sodium ions for membrane transport and also maintains the osmotic equilibrium of the medium. Agar acts as a solidifying agent.

Type of specimen

Recombinant strains of E.coli

Specimen Collection and Handling

For recombinant strain samples follow appropriate techniques for handling specimens as per established guidelines (3,4,5). 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. Some strains may show poor growth due to nutritional variations.
  2. Further biochemical and serological tests must be carried out for complete 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 yellow coloured granulated media

Gelling
Firm, comparable with 1.5% Agar gel

Colour and Clarity of prepared medium
Yellow to amber coloured, clear to slightly opalescent gel forms in Petri plates

Quantity
Each capsule contains 7.63 grams of medium sufficient for 250 ml media

Reaction
Reaction of 3.05% w/v aqueous solution at 25°C. pH: 7.0±0.2

pH
6.80-7.20

Cultural Response
Cultural characteristics observed after an incubation of 18-24 hours at 35-37°C with added 2.5 ml of 20% dextrose solution to 250 ml of EC1726CCL

Organism Growth Inoculum
(CFU)
Recovery
Escherichia coli ATCC 23724 luxuriant 50-100 >=70%
Escherichia coli ATCC 25922 (00013*) luxuriant 50-100 >=70%
Escherichia coli DH5 alpha MTCC 1652 luxuriant 50-100 >=70%

Key: (*) Corresponding WDCM numbers

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 (1,2).

More Information
Product Name HiEncap™ Luria Agar Base, Miller's Modification
SKU EC1726CCL
Product Type HiEncap™
Physical Form Granulated Media in Gelatin Capsule
Origin Animal
Packaging type HDPE Jar
References 1. Miller ,J.H. 1772.Experiments in molecular genetics. Cold spring Harbor Laboratory,Cold spring Harbor, New York.
2.Sambrook, J., E.F. Fritsch and T. Maniatis. 1989.Molecular cloning: A laboratory manual, 2nd ed., Cold Spring Harbor Laboartory, Cold Spring Harbor, New York.
3.Lennox E.S. 1955, Transduction of Linked Genetic Characters of the host by bacteriophage P1., Virology, 1:190
Customized Product Available No
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