Measuring Mutation Rates Using the Luria-Delbrück Fluctuation Assay

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1672)

Abstract

The Luria-Delbrück fluctuation assay is one of the most commonly used methods for measuring the mutation rate in microorganisms. Specifically, it is used to measure the mutation rate at a particular locus or loci at which mutations give rise to a selectable phenotype. Here, I outline the essential features of performing Luria-Delbrück fluctuation assays as well as common missteps and tips for improving the accuracy of mutation rate estimates. In addition, I provide tools for analyzing data from fluctuation assays. This 96-well plate protocol has been optimized for use in yeast but should perform equally well for a range of microorganisms using standard microbiological methods.

Key words

Mutation rate Fluctuation test Poisson distribution 

Notes

Acknowledgments

I thank Sean Buskirk, Katie Fisher, and Dan Marad for comments on this manuscript. This work was supported by a New Investigator grant from the Charles E. Kaufman Foundation of The Pittsburgh Foundation.

Supplementary material

393503_1_En_3_MOESM1_ESM.docx (64 kb)
LangFluctuationAssay-161201(DOCX 64 kb).

References

  1. 1.
    Luria S, Delbrück M (1943) Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28:491–511PubMedPubMedCentralGoogle Scholar
  2. 2.
    Ma WT, Sandri GH, Sarkar S (1992) Analysis of the Luria-Delbrück distribution using discrete convolution powers. J App Prob 29:255–267Google Scholar
  3. 3.
    Stewart FM (1994) Fluctuation tests: how reliable are the estimates of mutation rates? Genetics 137(4):1139–1146PubMedPubMedCentralGoogle Scholar
  4. 4.
    Rosche WA, Foster PL (2000) Determining mutation rates in bacterial populations. Methods 20(1):4–17CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Foster PL (2006) Methods for determining spontaneous mutation rates. Methods Enzymol 409:195–213CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Sarkar S, Ma WT, Sandri GH (1992) On fluctuation analysis: a new, simple and efficient method for computing the expected number of mutants. Genetica 85(2):173–179CrossRefPubMedGoogle Scholar
  7. 7.
    Lang GI, Murray AW (2008) Estimating the per-base-pair mutation rate in the yeast Saccharomyces cerevisiae. Genetics 178(1):67–82CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Gillet-Markowska A, Louvel G, Fischer G (2015) bz-rates: a web tool to estimate mutation rates from fluctuation analysis. G3 (Bethesda) 5(11):2323–2327CrossRefGoogle Scholar
  9. 9.
    Hall BM et al (2009) Fluctuation analysis CalculatOR: a web tool for the determination of mutation rate using Luria-Delbrück fluctuation analysis. Bioinformatics 25(12):1564–1565CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Stewart FM (1991) Fluctuation analysis: the effect of plating efficiency. Genetica 84(1):51–55CrossRefPubMedGoogle Scholar
  11. 11.
    Stewart FM, Gordon DM, Levin BR (1990) Fluctuation analysis: the probability distribution of the number of mutants under different conditions. Genetics 124(1):175–185PubMedPubMedCentralGoogle Scholar
  12. 12.
    Zheng Q (2005) New algorithms for Luria-Delbrück fluctuation analysis. Math Biosci 196(2):198–214CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Department of Biological SciencesLehigh UniversityBethlehemUSA

Personalised recommendations