Quick Analysis of Sedimentary Ancient DNA Using quicksand

Abstract Ancient DNA extracted from the sediments of archaeological sites (sedaDNA) can provide fine-grained information about the composition of past ecosystems and human site use, even in the absence of visible remains. However, the growing amount of available sequencing data and the nature of the data obtained from archaeological sediments pose several computational challenges; among these, the rapid and accurate taxonomic classification of sequences. While alignment-based taxonomic classifiers remain the standard in sedaDNA analysis pipelines, they are too computationally expensive for the processing of large numbers of sedaDNA sequences. In contrast, alignment-free methods offer fast classification but suffer from higher false-positive rates. To address these limits, we developed quicksand, an open-source Nextflow pipeline designed for rapid and accurate taxonomic classification of mammalian mitochondrial DNA in sedaDNA samples. quicksand combines fast alignment-free classification using KrakenUniq with post-classification mapping, filtering, and ancient DNA authentication. Based on simulations and reanalyses of published datasets, we demonstrate that quicksand achieves accuracy and sensitivity comparable to or better than existing methods, while significantly reducing runtime. quicksand offers an easy workflow for large-scale screening of sedaDNA samples for archaeological research and is freely available at https://github.com/mpieva/quicksand.