Canine melanoma is one of the most common oral cancers seen in dogs, but can also affect other mucocutaneous areas such as the lips and eyes. These masses can be pigmented or not (amelanotic) and patients diagnosed with this disease have varying survival rates. Treatment typically consists of excision of the tumor and surrounding area, along with radiation or chemotherapy, and routine vaccines to prevent residual tumor cells from developing. This project aims to determine the heterogeneity of canine melanoma cell lines through performing single-cell transcriptome (scRNA-seq) analysis. ScRNA-seq provides a benefit over bulk sequencing of separating the transcript reads cell-by-cell, allowing for individual cells to be sequenced and cell populations identified. Analysis of such results can be used to cluster characteristically similar cells within a cell line. Through bioinformatic analysis of the data and comparison to a canine reference genome, novel biomarkers and pathways along with up and downregulation of genes will be determined, which can potentially serve as targets in the development of future therapies. Finally, by comparing the results of three distinct cell lines coming from very different clinical cases, the heterogeneity between tumor types, such as that of melanotic versus amelanotic, can be studied.