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Odonata comprise approximately 6400 species with extensive morphological and ecological diversity, specifically their colour variation, flight behaviour patterns, and breadth of ecological niches. Additionally, their phylogenetic placement within Insecta as descendants of the first winged insects make them ideal candidates for exploring evolutionary forces that have shaped diversity patterns (e.g., diversification rate shifts) as well as character evolution (e.g., flight behaviour, colour). Even though morphological and ecological traits are relatively well known for most of odonate taxa, the lack of well-supported phylogenetic hypothesis across Odonata have limited the capability of evaluating evolutionary phenomena in a comparative context. Previous studies using various taxon sampling schemes and data types (i.e. morphology, targeted locus approaches) to reconstruct odonate relationships failed to resolve several interfamilial relationships, specifically in groups with likely incomplete lineage sorting and/or introgression. Even though a recent study by Bybee et al. (2021) incorporated genomic-scale anchored hybrid enrichment (AHE) data for phylogenetic reconstruction, the relatively limited taxon sampling likely precluded resolution within the problematic groups. Our study, also targeting AHE loci, greatly expand taxon odonate genera, which resulted in 729 newly generated samples in a addition to 142 samples from Bybee et al. (2021) for a total of 831. With around 500 AHE loci, we aim to resolve historically difficult relationships and construct a robust ordinal phylogeny of Odonata, which will be used as the evolutionary framework to clarify taxonomic classifications and test evolutionary hypotheses regarding shifts in flight behaviours, colours, and diversification rates. 

Abstract The use of gDNAs isolated from museum specimens for high throughput sequencing, especially targeted sequencing in the context of phylogenetics, is a common practice. Yet, little understanding has been focused on comparing the quality of DNA and results of sequencing museum DNAs. Dragonflies and damselflies are ubiquitous in freshwater ecosystems and are commonly collected and preserved insects in museum collections hence their use in this study. However, the history of odonate preservation across time and museums has resulted in wide variability in the success of viable DNA extraction, necessitating an assessment of their usefulness in genetic studies. Using Anchored Hybrid Enrichment probes, we sequenced DNA from samples at 2 museums, 48 from the American Museum of Natural History (AMNH) in NYC, USA and 46 from the Naturalis Biodiversity Center (RMNH) in Leiden, Netherlands ranging from global collection localities and across a 120-year time span. We recovered at least 4 loci out of an >1,000 locus probe set for all samples, with the average capture being ~385 loci (539 loci on average when a clade of ambiguous taxa omitted). Neither specimen age nor size was a good predictor of locus capture, but recapture rates differed significantly between museums. Samples from the AMNH had lower overall locus capture than the RMNH, perhaps due to differences in specimen storage over time.