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    This dataset is derived from a comparative study evaluating six DNA extraction methods for their efficiency in recovering diatom sedimentary ancient DNA (sedaDNA) from Antarctic marine sediments. Sediment samples were collected from two sites: U1536C (Scotia Sea, West Antarctica) and KC02 (Sabrina Coast, Totten Glacier Region, East Antarctica). Each of the six extraction methods was applied to the same set of samples. Following shotgun metagenomic sequencing, the methods were assessed based on metrics such as diatom DNA recovery, average fragment length, and taxonomic diversity. The purpose of the study was to identify the optimal extraction approach for maximizing the yield and quality of diatom sedaDNA, thereby improving its utility for paleoenvironmental reconstruction.