G3: Genes|Genomes|Genetics Early Online, published on June 29, 2020 as doi:10.1534/g3.120.401028 1 Measuring genome sizes using read-depth, k-mers, and flow 2 cytometry: methodological comparisons in beetles (Coleoptera) 3 James M. Pflug,*, Valerie Renee Holmes,† Crystal Burrus,† J. Spencer Johnston,† and David R. 4 Maddison* 5 6 *Department of Integrative Biology, Oregon State University, Corvallis, OR 97331, USA 7 †Department of Entomology, Texas A&M University, College Station, TX 77843, USA 8 1 Corresponding author: 3029 Cordley Hall, Department of Integrative Biology, Oregon State University, Corvallis, OR 97331 USA. E-mail:
[email protected] © The Author(s) 2020. Published by the Genetics Society of America. 2 9 Abstract 10 Measuring genome size across different species can yield important insights into 11 evolution of the genome and allow for more informed decisions when designing next-generation 12 genomic sequencing projects. New techniques for estimating genome size using shallow 13 genomic sequence data have emerged which have the potential to augment our knowledge of 14 genome sizes, yet these methods have only been used in a limited number of empirical studies. In 15 this project, we compare estimation methods using next-generation sequencing (k-mer methods 16 and average read depth of single-copy genes) to measurements from flow cytometry, a standard 17 method for genome size measures, using ground beetles (Carabidae) and other members of the 18 beetle suborder Adephaga as our test system. We also present a new protocol for using read- 19 depth of single-copy genes to estimate genome size.