Please refer disclaimer Overleaf.

Luria Broth Base, Miller's Modification M1725Intended UseRecommended for the cultivation and maintenance of recombinant strains of Escherichia coli with or without addition of glucose.Composition**

Ingredients Gms / LitreTryptone 10.000Yeast extract 5.000Sodium Chloride 0.500Final pH ( at 25°C) 7.0±0.2**Formula adjusted, standardized to suit performance parameters

DirectionsSuspend 15.5 grams in 1000 ml purified / distilled water. Heat if necessary 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 10 ml of 20% glucose solution. Mix thoroughly and pour into sterile test tubes or flasks.

Principle And Interpretation

This medium is based on original Luria broth formula described by Miller for the growth and maintenance of E.coli strains used in molecular microbiology (4). Luria broth 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 Broth 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.

Type of specimen Recombinant strains of E.coli

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.

Specimen Collection and Handling

Warning and PrecautionsRead 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

expiry period Performance and EvaluationPerformance of the medium is expected when used as per the direction on the label within thewhen stored at recommended temperature.

1. Some strains may show poor growth due to nutritional variations.

2. Further biochemical and serological tests must be carried out for complete identification.

HiMedia Laboratories Technical Data

Disclaimer :

User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained inthis and other related HiMedia™ publications. The information contained in this publication is based on our research and developmentwork and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes tospecifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use butfor laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should notbe considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.

Escherichia coli DH5 alphaMTCC 1652

luxuriant 50-100

Reference

Revision : 02 / 2019

Quality ControlAppearanceCream to yellow homogeneous free flowing powderColour and Clarity of Prepared MediumYellow to amber coloured clear to slightly opalescent solutionReactionReaction of 1.55% w/v aqueous solution at 25°C. pH : 7.0±0.2pH6.80-7.20Cultural ResponseCultural characteristics observed after an incubation at 35-37°C for 18-24 hours .Organism Growth Inoculum

(CFU)

Escherichia coli ATCC23724

luxuriant 50-100

Escherichia coli ATCC25922 (00013*)

luxuriant 50-100

Key : (*) Corresponding WDCM numbers.

Storage and Shelf LifeStore 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.

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).

Disposal

4. Miller ,J.H. 1772.Experiments in molecular genetics. Cold spring Harbor Laboratory Cold spring Harbor, New York.5. Sambrook, J., E.F. Fritsch and T. Maniatis. 1989.Molecular cloning: A laboratory manual, 2nd ed., Cold Spring HarborLaboartory, Cold Spring Harbor, New York.

3. Lennox E.S. 1955, Transduction of Linked Genetic Characters of the host by bacteriophage P1., Virology, 1:190.

2. Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1.

1. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.

HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate office : A-516,Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147 1919 Email: techhelp@himedialabs.com Website: www.himedialabs.com