-The NExt-generation sequencing Analysis Toolkit (NEAT) is an open-source Python package that creates simulated next-generation sequencing datasets. NEAT’s simulations account for a wide range of sequencing parameters (e.g., DNA read fragment length, sequencing error rates, mutation frequencies, etc.) and allow users to customize their sequencing data.<sup>1</sup> Since the original release of NEAT in 2016, most scripts have been greatly modified, and NEAT is currently on version 4.2. The code has undergone significant ongoing changes since 2020. Upgrading to Python 3 has enabled NEAT to achieve a flexible and intuitive user interface with minimal dependencies. The toolkit is optimized for both speed and accuracy, and new features have been implemented, such as improved ploidy simulation, mutation modeling, and the ability to model mutational profiles directly from data. A summary of algorithmic changes is provided in Table 1.
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