4.4 Article

Sizing up arthropod genomes: an evaluation of the impact of environmental variation on genome size estimates by flow cytometry and the use of qPCR as a method of estimation

Journal

GENOME
Volume 56, Issue 9, Pages 505-510

Publisher

CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
DOI: 10.1139/gen-2013-0044

Keywords

Arthropoda; genome size; flow cytometry; real-time PCR; polymerase chain reaction

Funding

  1. NSERC
  2. Canada Research Chairs Program
  3. Canada Foundation for Innovation
  4. British Columbia Knowledge Development Fund
  5. Genome Canada
  6. Genome British Columbia
  7. Genome Alberta

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A study was undertaken to evaluate both a pre-existing method and a newly proposed approach for the estimation of nuclear genome sizes in arthropods. First, concerns regarding the reliability of the well-established method of flow cytometry relating to impacts of rearing conditions on genome size estimates were examined. Contrary to previous reports, a more carefully controlled test found negligible environmental effects on genome size estimates in the fly Drosophila melanogaster. Second, a more recently touted method based on quantitative real-time PCR (qPCR) was examined in terms of ease of use, efficiency, and (most importantly) accuracy using four test species: the flies Drosophila melanogaster and Musca domestica and the beetles Tribolium castaneum and Dendroctonus ponderosa. The results of this analysis demonstrated that qPCR has the tendency to produce substantially different genome size estimates from other established techniques while also being far less efficient than existing methods.

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