Genome size—the total haploid content of nuclear DNA— is constant in all cells in individuals within a species, but differs among species. Consequently, the genome size is a quantifiable genetic signature that not only characterizes a species, but it can reflect chromatin modifications, which play fundamental roles in most biological processes that are involved in the manipulation and expression of DNA. This characteristic makes the genome size a crucial parameter for genetic research on endemic aquatic species and for genetic manipulations in aquaculture species. Technologies for genetic assessments and improvements applied to fishery and aquaculture species use genome size values as a means by which hybrids, polyploids, and sex can be identified, when sex chromosomes exist. The objectives of this study were to determine genome sizes of aquatic species with economic and biological importance along the Pacific coast of Mexico, as well as to identify the appropriate reference standards for use in this study. Blood, hemolymph or milt were collected from 10 species occurring along the coast of Baja California: Sablefish Anoplopoma fimbria, Black Snapper Lutjanus novemfasciatus, California Halibut Paralichthys californicus, Pacific Sardine Sardinops sagax, Flag Rockfish Sebastes rubrivinctus, Starry Rockfish Sebastes constellatus, Totoaba Totoaba macdonaldi, Whiteleg Shrimp Litopenaeus vannamei and two Yellowtail Seriola lalandi and S. dorsalis. Nuclear DNA was stained with propidium iodide solution and the genome size was determined by flow cytometry, with results ranging from 0.61 pg (1.22 pg/diploid cell) to 2.59 pg (5.18 pg/diploid cell), with the smallest value in Sablefish and the largest in the Whiteleg Shrimp. No significant differences were detected (P ≤ 0.05) among individuals of the same species; the likely reason behind any dissimilar DNA content values with those from the literature were differences in methodologies or variations in genetics. Red-ear Slider Turtle Trachemys scripta elegans 2.65 pg (5.30 pg/diploid cell) and Red Junglefowl Gallus gallus 1.27 pg (2.54 pg/diploid cell) were chosen as the standards for reference values. These results establish the basis for the Mexican National Aquatic Genetic Resources project supporting genetic improvements for aquaculture and conservation status parameters for fisheries species.