title: 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.
abstract: 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.
authors.affiliation: a Department of Integrative Biology, University of Guelph, 50 Stone Road E., Guelph, ON N1G 2W1, Canada.
authors.forename: T Ryan
authors.initials: TR
authors.lastname: Gregory
authors.forename: Paula
authors.initials: P
authors.lastname: Nathwani
authors.forename: Tiffany R
authors.initials: TR
authors.lastname: Bonnett
authors.forename: Dezene P W
authors.initials: DP
authors.lastname: Huber
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005434:Flow Cytometry / Q000379:methods*; D059646:Genome Size; D049750:Genome, Insect; D006793:Houseflies / Q000235:genetics*; D006813:Humidity; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013696:Temperature; D014231:Tribolium / Q000235:genetics