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Related Experiment Videos

Large-scale production of lambda bacteriophage and purified lambda deoxyribonucleic acid.

A De Czekala, D Luk, P Bartl

    Applied Microbiology
    |April 1, 1972
    PubMed
    Summary

    Researchers developed a standardized method to produce large quantities of heat-inducible phage lambda mutant (lambdaCI857). This process involves polyethylene glycol precipitation, cesium chloride density gradient banding, and cold phenol extraction for purified, unbroken lambda DNA molecules.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Biochemistry

    Background:

    • Phage lambda is a bacteriophage widely used in molecular biology.
    • Efficient production of phage lambda mutants is crucial for various research applications.
    • LambdaCI857 is a heat-inducible mutant of phage lambda.

    Purpose of the Study:

    • To establish a standardized protocol for obtaining large quantities of phage lambda mutant (lambdaCI857).
    • To outline a purification strategy for both the phage and its deoxyribonucleic acid (DNA).

    Main Methods:

    • Phage lambda mutant (lambdaCI857) induction and harvesting.
    • Polyethylene glycol (C 6000) precipitation for phage concentration.
    • Cesium chloride (CsCl) density gradient banding for phage purification.

    Related Experiment Videos

  • Cold phenol extraction for deoxyribonucleic acid (DNA) isolation.
  • Sucrose density gradient sedimentation for DNA purification.

    • Main Results:

    • Successfully obtained large amounts of heat-inducible phage lambda mutant (lambdaCI857) under standardized conditions.
    • Phage was efficiently harvested using polyethylene glycol precipitation.
    • Purified phage DNA yielded a homogenous population of unbroken lambda DNA molecules.

    Conclusions:

    • The described method provides a reliable and scalable approach for producing high-quality phage lambda mutant (lambdaCI857) and its DNA.
    • This protocol facilitates downstream applications requiring pure, intact lambda DNA.
    • Standardized conditions ensure reproducibility in phage lambda mutant production.