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Polyploidy is widespread across the tree of life, yet there remain large gaps in our understanding of polyploid evolution. After their formation, the genome and gene expression architecture of polyploids may undergo rapid changes. What is unclear is how independent changes in isolated populations may affect the evolution of a polyploid species when these populations come into contact. In this study, we aimed to elucidate the area of contact between two tetraploid lineages of the gray treefrog (Hyla versicolor). Although previous research provided a rough estimate of where these different lineages co-occur, we lack information about the extent of these lineage across large geographic regions and about the position of possible contact zones. In preliminary work, we sequenced a mitochondrial marker (containing fixed differences between lineages) along a transect spanning the putative contact region, to identify the exact zone where the lineages co-occur. Despite fine-scale sampling along a transect with multiple populations, we were unable to identify sites where the mitochondrial lineages co-occurred. This Honors in the Major project analyzed additional samples from the Northeast region of the United States. In this project, I aimed to locate populations of H. versicolor, determine their lineages, and characterize acoustic differences between previously and recently collected samples to further compare the characteristics of both lineages.