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Plating Bacteriophage M13

Michael R Green, Joseph Sambrook

    Cold Spring Harbor Protocols
    |October 5, 2017
    PubMed

    Insights

    Bacteriophage M13 forms plaques by infecting bacteria in semisolid media. Infected cells grow slower, creating visible plaques on a faster-growing bacterial lawn within hours.

    Area of Science:

    • Microbiology
    • Virology
    • Molecular Biology

    Background:

    • Bacteriophage M13 is a virus that infects bacteria.
    • Phage plaques are visible areas formed by viral activity in a bacterial lawn.
    • Understanding plaque formation is crucial for viral stock titration and study.

    Purpose of the Study:

    • To describe a protocol for plating bacteriophage M13 stocks.
    • To detail the conditions for plaque visualization.
    • To explain the biological basis of M13 plaque formation.

    Main Methods:

    • Infecting single Escherichia coli cells with bacteriophage M13.
    • Culturing infected bacteria in semisolid medium (agar or agarose).
    • Incubating plates at 37°C for plaque development.

    Main Results:

    • Bacteriophage M13 plaques are formed by slower-growing infected cells.
    • Limited diffusion of progeny viruses in semisolid media facilitates plaque formation.
    • Plaques are detectable within 4-8 hours of incubation.

    Conclusions:

    • The described protocol allows for reliable plating and visualization of bacteriophage M13 plaques.
    • M13 plaques serve as indicators of viral infection and replication.
    • This method is essential for quantifying bacteriophage M13 stocks.

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