{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T19:04:06Z","timestamp":1791486246765,"version":"4.3.4"},"reference-count":24,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,9]]},"abstract":"Abstract<\/jats:title>\n \n Phages remain severely under\u2010sampled, largely due to the limitations of current isolation methods, which are labor\u2010intensive, low\u2010throughput, and often restricted to a narrow set of bacterial hosts. To address this gap, we developed a robust and scalable field\u2010to\u2010genome workflow that leverages laboratory automation to screen environmental samples against a comprehensive 188\u2010strain bacterial host library using a 384\u2010well plate format. We demonstrate the workflow using\n Acinetobacter baumannii<\/jats:italic>\n as the primary model host. Phage activity is detected through Bacterial Lysis Observation over Time assay (BLOTS) using spectrophotometry. Subsequently, isolation and subculture of the phages are done with a single layer agar spot assay. Phage genomes are then sequenced using long\u2010read technology and annotated. This enables systematic and reproducible phage detection and enrichment from diverse sample types. The use of large numbers of host strains streamlines phage discovery and substantially increases the chances of isolating novel phages. The BLOTS high\u2010throughput approach can also be used for screening phage collections against specified clinical hosts of interest for phage research and candidate selection for phage therapy applications. \u00a9 2026 The Author(s).\n Current Protocols<\/jats:italic>\n published by Wiley Periodicals LLC.\n <\/jats:p>\n \n Basic Protocol 1<\/jats:bold>\n : Preparation of host libraries and multi\u2010host primary enrichment\n <\/jats:p>\n \n Basic Protocol 2<\/jats:bold>\n : Preparation of the BLOTS screening plate using an automated liquid handler\n <\/jats:p>\n \n Alternate Protocol 1<\/jats:bold>\n : Manual phage screening without laboratory automation\n <\/jats:p>\n \n Basic Protocol 3<\/jats:bold>\n : Phage screening using BLOTS\n <\/jats:p>\n \n Basic Protocol 4<\/jats:bold>\n : Phage validation using the single\u2010layer agar spot assay\n <\/jats:p>\n \n Basic Protocol 5<\/jats:bold>\n : Phage amplification for downstream applications\n <\/jats:p>\n \n Basic Protocol 6<\/jats:bold>\n : Phage concentration using PEG\u20108000 precipitation\n <\/jats:p>\n \n Alternate Protocol 2<\/jats:bold>\n : Phage concentration using centrifugal filtration\n <\/jats:p>\n \n Basic Protocol 7<\/jats:bold>\n : Automated DNA extraction with magnetic beads\n <\/jats:p>\n \n Basic Protocol 8<\/jats:bold>\n : Whole\u2010genome sequencing using long reads\n <\/jats:p>\n \n Basic Protocol 9<\/jats:bold>\n : De novo genome assembly using the Phage Galaxy platform\n <\/jats:p>\n \n Basic Protocol 10<\/jats:bold>\n : Annotation and characterization of phage genome\n <\/jats:p>\n \n Basic Protocol 11<\/jats:bold>\n : Host range testing and characterization of isolated phages using BLOTS\n <\/jats:p>","DOI":"10.1002\/cpz1.70454","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T16:45:45Z","timestamp":1788194745000},"source":"Crossref","is-referenced-by-count":1,"title":["Bacterial Lysis Observation Over Time Assay (BLOTS) for High\u2010Throughput Isolation, Sequencing, and Characterization of Bacteriophages"],"prefix":"10.1002","volume":"6","author":[{"given":"Sean Jia Le","family":"Pang","sequence":"first","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science, Interdisciplinary Graduate Programme Nanyang Technological University Singapore"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soon Keong","family":"Wee","sequence":"additional","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science Nanyang Technological University Singapore"},{"name":"These authors contributed equally to this work"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bobby Guo Wei","family":"Lee","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine Nanyang Technological University Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Odelia","family":"Yap","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine Nanyang Technological University Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric Peng Huat","family":"Yap","sequence":"additional","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science Nanyang Technological University Singapore"},{"name":"Lee Kong Chian School of Medicine Nanyang Technological University Singapore"},{"name":"National Centre for Infectious Diseases Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"e_1_2_18_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.594868"},{"key":"e_1_2_18_3_1","doi-asserted-by":"publisher","DOI":"10.5962\/bhl.title.6966"},{"key":"e_1_2_18_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43705\u2010023\u201000299\u20105"},{"key":"e_1_2_18_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010024\u201049710\u20102"},{"key":"e_1_2_18_6_1","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.2261"},{"key":"e_1_2_18_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.resmic.2008.05.001"},{"key":"e_1_2_18_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_18_9_1","doi-asserted-by":"publisher","DOI":"10.1055\/s\u20100034\u20101398388"},{"key":"e_1_2_18_10_1","first-page":"652","article-title":"Des relations num\u00e9riques entre bact\u00e9ries 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