PIMENTO: a primer inference toolkit to facilitate large-scale calling of amplicon sequence variants

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 the PrIMER infereNce TOolkit (PIMENTO), a Python package that 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.