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Measuring Microbial Mutation Rates with the Fluctuation Assay
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Measuring Microbial Mutation Rates with the Fluctuation Assay.

Rok Krašovec1, Huw Richards2, Guillaume Gomez3

  • 1School of Biological Sciences, The University of Manchester; rok.krasovec@manchester.ac.uk.

Journal of Visualized Experiments : Jove
|December 17, 2019
PubMed
Summary
This summary is machine-generated.

Fluctuation assays estimate microbial mutation rates using parallel cultures and selective markers. This method is cost-effective, less laborious than alternatives, and adaptable for various organisms and environments.

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Area of Science:

  • Microbiology
  • Genetics
  • Evolutionary Biology

Background:

  • Fluctuation assays are standard for estimating microbial mutation rates in liquid cultures.
  • The method involves inoculating parallel cultures, allowing growth, and then assessing resistant mutants on selective media.
  • Estimating mutation rates is crucial for understanding antimicrobial resistance, aging, and evolution.

Purpose of the Study:

  • To provide a comprehensive overview of fluctuation assays for mutation rate estimation.
  • To highlight the critical aspects, advantages, and limitations of the assay.
  • To discuss recent advancements addressing assay assumptions and expanding its applicability.

Main Methods:

  • Inoculating parallel cultures with a known number of sensitive cells.
  • Growing cultures for many generations in a non-selective environment.
  • Plating a subset of cultures on selective agar to count resistant mutants and estimating population size (Nt).
  • Using mathematical models to determine the number of mutational events (m) and calculate mutation rate (m/Nt).

Main Results:

  • Fluctuation assays provide a cost-effective and less laborious method for mutation rate estimation compared to alternatives.
  • The assay is adaptable to various organisms, genotypes, and environmental contexts, including microbial communities.
  • Recent analytical tools and theoretical studies enhance the accuracy and address limitations related to marker fitness and cell death.

Conclusions:

  • Fluctuation assays are a versatile and valuable tool for estimating spontaneous mutation rates in diverse biological systems.
  • The assay's adaptability and recent analytical improvements make it suitable for studying the impact of environmental factors on mutation rates.
  • Understanding mutation rates is fundamental for fields ranging from medicine (antimicrobial resistance, carcinogenesis) to evolutionary biology.