{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T18:11:13Z","timestamp":1789755073737,"version":"4.0.1"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100017700","name":"Henan Provincial Science and Technology Research Project","doi-asserted-by":"publisher","award":["222102310398"],"award-info":[{"award-number":["222102310398"]}],"id":[{"id":"10.13039\/501100017700","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013066","name":"Key Scientific Research Project of Colleges and Universities in Henan Province","doi-asserted-by":"publisher","award":["24A310002"],"award-info":[{"award-number":["24A310002"]}],"id":[{"id":"10.13039\/501100013066","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    The zebrafish (\n                    <jats:italic>Danio rerio<\/jats:italic>\n                    ) is a widely used vertebrate model in biomedical research, yet the rapid and cost-effective extraction of genomic DNA (gDNA) from adult zebrafish tissue remains a practical challenge for routine molecular applications. To address this, we systematically optimized and evaluated a Chelex-100\/proteinase K-based extraction protocol across eight adult zebrafish tissues (muscle, liver, heart, spleen, kidney, intestine, brain, and eggs) and compared its performance against that of a commercial kit. The optimized protocol, involving incubation at 56 \u00b0C for 15 min and boiling at 100 \u00b0C for 10 min, required approximately 25 min per sample. Across all tested tissues (3 biological replicates per tissue), the Chelex-100 method yielded gDNA concentrations ranging from 177.2\u2009\u00b1\u20093.5 ng\/\u00b5L to 674.5\u2009\u00b1\u200922.1 ng\/\u00b5L per 5 mg of tissue, which were generally higher than those obtained with the commercial kit. However, the purity (A260\/A280 ratios) was consistently lower for the Chelex-100 method (1.36\u20131.83) than for the kit (1.84\u20132.02), indicating the coextraction of proteinaceous impurities. Despite this, Chelex-extracted gDNA supported successful PCR amplification and Sanger sequencing of target fragments, with consistent amplicon quality. In real-time PCR assays, both methods produced amplification curves, but the Chelex-100 method yielded systematically higher Ct values, suggesting reduced sensitivity with less-pure templates. Crucially, the Chelex-100 method avoided hazardous organic solvents and required no column purification, with an estimated reagent cost of &lt;$0.20 per sample versus approximately $2.00 per sample for the commercial kit. We conclude that this optimized Chelex-100\/proteinase K protocol provides a rapid, economical, and operationally simple alternative for routine gDNA extraction from adult zebrafish tissues and is particularly suitable for PCR-based genotyping, sequencing and qPCR applications, though its lower purity warrants careful consideration for high-sensitivity quantitative assays.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0358591","type":"journal-article","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T17:50:35Z","timestamp":1789753835000},"page":"e0358591","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["A rapid and cost-effective Chelex-100 resin-based method for genomic DNA extraction from adult zebrafish tissues"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3061-5359","authenticated-orcid":true,"given":"Wei","family":"Feng","sequence":"first","affiliation":[{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["School of Basic Medicine and Forensic Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Institute of Special Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Forensic Center"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boya","family":"Li","sequence":"additional","affiliation":[{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["School of Basic Medicine and Forensic Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiqiong","family":"Niu","sequence":"additional","affiliation":[{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["School of Basic Medicine and Forensic Medicine"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiannan","family":"Xu","sequence":"additional","affiliation":[{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["School of Basic Medicine and Forensic Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Institute of Special Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Forensic Center"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiandun","family":"Zhai","sequence":"additional","affiliation":[{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["School of Basic Medicine and Forensic Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Institute of Special Medicine"]},{"name":"Henan University of Science and Technology","place":["Luoyang, China"],"department":["Forensic Center"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,18]]},"reference":[{"issue":"1","key":"pone.0358591.ref001","doi-asserted-by":"crossref","first-page":"194","DOI":"10.3390\/ijms25010194","article-title":"Zebrafish (Danio rerio) as a model for the study of developmental and cardiovascular toxicity of electronic cigarettes","volume":"25","author":"E Hussen","year":"2023","journal-title":"Int J Mol Sci"},{"key":"pone.0358591.ref002","doi-asserted-by":"crossref","first-page":"177776","DOI":"10.1016\/j.ejphar.2025.177776","article-title":"Towards zebrafish model applications in drug discovery targeting central nervous system diseases and neurotoxicity assessment","volume":"1002","author":"H Li","year":"2025","journal-title":"Eur J 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