PIMENTO: A PrIMEr infereNce TOolkit to facilitate large-scale calling of amplicon sequence variants

Abstract The identification of amplicon sequence variants from DNA metabarcoding data is a common method for revealing the taxonomic makeup of environmental samples, and for allowing comparative studies between similar datasets. A significant hurdle to the large-scale calling of amplicon sequence variants from publicly available nucleotide datasets is the heterogeneous presence of primer sequences in reads, the removal of which is a necessary pre-processing step for this form of analysis. Furthermore, as the details of the experimental primers are rarely captured in the metadata associated with the sequence records, there is a need for a method that can automatically infer the presence and identity of primers in sequencing data. In this work, we introduce PIMENTO, a Python package which uses a dual-strategy approach for identifying primers that are present in sequencing reads to enable their removal, and therefore facilitate amplicon sequence variant calling at scale. Competing Interest Statement The authors have declared no competing interest. Footnotes Have added an extra snippet in the funding acknowledgements, and added a missing bit in the package availability section.