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However, current protocols require large DNA input and are cost-prohibitive, which limits their application to phage collections that typically are large and have low biomass. In order to address this, we have developed\n                    <jats:italic>Plaque-2-seq<\/jats:italic>\n                    , a robust and cost-effective workflow for high-throughput phage genome sequencing that will transform the speed and cost of attaining phage genomes.\n                    <jats:italic>Plaque-2-seq<\/jats:italic>\n                    combines low-input transposase-based library preparation, amplification, nanopore sequencing and optimized assembly steps tailored to phage genomes. We applied the method to phages isolated on seven genetically diverse bacterial hosts:\n                    <jats:italic>Escherichia<\/jats:italic>\n                    ,\n                    <jats:italic>Pseudomonas<\/jats:italic>\n                    ,\n                    <jats:italic>Synechococcus<\/jats:italic>\n                    ,\n                    <jats:italic>Enterococcus<\/jats:italic>\n                    ,\n                    <jats:italic>Klebsiella, Serratia<\/jats:italic>\n                    and\n                    <jats:italic>Enterobacter<\/jats:italic>\n                    . High-quality genome assemblies were validated using CheckV and benchmarked against previously sequenced phage isolates. Compared to standard Illumina sequencing,\n                    <jats:italic>Plaque-2-seq<\/jats:italic>\n                    offers up to ~5\u201310-fold savings in sequencing price for individual labs. Furthermore, it substantially decreases the time required to produce a phage genome, once a plaque is obtained. Offering the ability to routinely obtain hundreds of phage genome sequences a week, with minimal hands-on time.\n                    <jats:italic>Plaque-2-seq<\/jats:italic>\n                    enables systematic genomic characterization of phage isolates, facilitating taxonomic classification, for the development of large-scale phage biobanks.\n                  <\/jats:p>","DOI":"10.1099\/mgen.0.001672","type":"journal-article","created":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T16:15:53Z","timestamp":1777047353000},"source":"Crossref","is-referenced-by-count":0,"title":["Rolling out Plaque-2-seq: a single plaque sequencing approach enabling rapid, low-cost sequencing of phages directly from plaques"],"prefix":"10.1099","volume":"12","author":[{"given":"Slawomir","family":"Michniewski","sequence":"first","affiliation":[{"name":"Becky Mayer Centre for Phage Research, University of Leicester, University Road, Leicester, 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