{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T18:35:52Z","timestamp":1762540552479,"version":"build-2065373602"},"reference-count":22,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T00:00:00Z","timestamp":1762473600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["INAU II 421733\/2017-9"],"award-info":[{"award-number":["INAU II 421733\/2017-9"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005286","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Mato Grosso","doi-asserted-by":"publisher","award":["PRO.000339\/2023"],"award-info":[{"award-number":["PRO.000339\/2023"]}],"id":[{"id":"10.13039\/501100005286","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["MCTIC\/CNPq 405706\/2022-7"],"award-info":[{"award-number":["MCTIC\/CNPq 405706\/2022-7"]}],"id":[{"id":"10.13039\/501100003593","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 present study demonstrates the viability of extracting DNA for genomic analyses from cell suspensions prepared for cytogenetic analyses, specifically, samples fixed in Carnoy\u2019s solution (3:1 methanol: acetic acid) and stored at \u201320 \u00b0C for more than 10 years. Cell suspensions from 27 specimens of fish of the subfamily Loricariinae, which had been prepared originally for cytogenetic studies, were used to test a DNA extraction procedure based on a routinely-used protocol with specific minor adjustments. The spectrophotometric and electrophoretic analyses of the extracted samples revealed DNA at good concentrations (5.9\u20131739 ng.\u03bcL\n                    <jats:sup>-1<\/jats:sup>\n                    ) and high purity (A260\/A280 ratios of 1.75\u20132.07), indicating negligible contamination and no significant fragmentation. The integrity of the material was confirmed by the successful amplification by PCR of four different genes (COI, 5S, 18S, and RAG2), with the COI gene being sequenced efficiently. These results demonstrate that DNA can be extracted from samples collected for other purposes, even after long-term storage, producing high-quality genomic DNA from sources that might otherwise be overlooked. The exploitation of these samples as a source of DNA represents a useful potential strategy for the acquisition of material for analysis when fresh samples are scarce or difficult to obtain. This novel approach expands considerably the possibilities for retrospective molecular studies, especially in the fields of conservation and systematics.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0335898","type":"journal-article","created":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T18:24:23Z","timestamp":1762539863000},"page":"e0335898","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Efficient DNA extraction from cytogenetic suspensions: A new possibility for obtaining DNA, with potential applications in studies of molecular markers"],"prefix":"10.1371","volume":"20","author":[{"given":"Ge\u00f3rgia Liz Monteiro","family":"Sant\u2019ana-Arruda","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paulo Cesar","family":"Venere","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2753-0613","authenticated-orcid":true,"given":"Daniela Cristina","family":"Ferreira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,11,7]]},"reference":[{"issue":"22","key":"pone.0335898.ref001","doi-asserted-by":"crossref","first-page":"4692","DOI":"10.1093\/nar\/25.22.4692","article-title":"Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques","volume":"25","author":"SM Aljanabi","year":"1997","journal-title":"Nucleic Acids Res"},{"issue":"1","key":"pone.0335898.ref002","doi-asserted-by":"crossref","first-page":"519","DOI":"10.4238\/vol10-1gmr1055","article-title":"A simplified universal genomic DNA extraction protocol suitable for PCR","volume":"10","author":"TY Wang","year":"2011","journal-title":"Genet Mol Res"},{"issue":"4","key":"pone.0335898.ref003","doi-asserted-by":"crossref","DOI":"10.1590\/S1516-84842004000400008","article-title":"Extra\u00e7\u00e3o de DNA de materiais de arquivo e fontes escassas para utiliza\u00e7\u00e3o em rea\u00e7\u00e3o de polimeriza\u00e7\u00e3o em cadeia (PCR)","volume":"26","author":"JA Barea","year":"2004","journal-title":"Rev Bras Hematol Hemoter"},{"issue":"4","key":"pone.0335898.ref004","doi-asserted-by":"crossref","DOI":"10.1590\/S1516-84842004000400001","article-title":"DNA e aperfei\u00e7oamento das t\u00e9cnicas de extra\u00e7\u00e3o","volume":"26","author":"V Bueno","year":"2004","journal-title":"Rev Bras Hematol Hemoter"},{"issue":"3","key":"pone.0335898.ref005","doi-asserted-by":"crossref","first-page":"379","DOI":"10.2307\/1223408","article-title":"Plant DNA isolation: a method to efficiently remove polyphenolics, polysaccharides, and RNA","volume":"44","author":"DV Jobes","year":"1995","journal-title":"TAXON"},{"issue":"2","key":"pone.0335898.ref006","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1007\/BF02774246","article-title":"An efficient protocol for genomic DNA extraction fromCitrus species","volume":"21","author":"Y-J Cheng","year":"2003","journal-title":"Plant Mol Biol Rep"},{"issue":"3","key":"pone.0335898.ref007","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1078\/1439-6092-00071","article-title":"DNA, PCR and formalinized animal tissue\u00a0? a short review and protocols","volume":"3","author":"C Schander","year":"2003","journal-title":"Organisms Diversity & Evolution"},{"issue":"5","key":"pone.0335898.ref008","article-title":"Sequencing degraded DNA from non-destructively sampled museum specimens for RAD-tagging and low-coverage shotgun phylogenetics","volume":"9","author":"MM-Y Tin","year":"2014","journal-title":"PLoS One"},{"key":"pone.0335898.ref009","first-page":"103","article-title":"Cytotaxonomic considerations on Hoplias lacerdae (Pisces, Erythrinidae)","volume":"1","author":"LAC Bertollo","year":"1978","journal-title":"Brazilian J Genetic"},{"issue":"9","key":"pone.0335898.ref010","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1007\/BF01923555","article-title":"A method for chromosome preparations from large fish specimens using in vitro short-term treatment with colchicine","volume":"49","author":"F Foresti","year":"1993","journal-title":"Experientia"},{"issue":"1462","key":"pone.0335898.ref011","doi-asserted-by":"crossref","first-page":"1847","DOI":"10.1098\/rstb.2005.1716","article-title":"DNA barcoding Australia\u2019s fish species","volume":"360","author":"RD Ward","year":"2005","journal-title":"Philos Trans R Soc Lond B Biol Sci"},{"key":"pone.0335898.ref012","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1186\/1471-2148-11-275","article-title":"Phylogenetic relationships within the speciose family Characidae (Teleostei: Ostariophysi: Characiformes) based on multilocus analysis and extensive ingroup sampling","volume":"11","author":"C Oliveira","year":"2011","journal-title":"BMC Evol Biol"},{"issue":"1","key":"pone.0335898.ref013","first-page":"324","article-title":"Phylogenetic relationships of new world needlefishes (Teleostei: Belonidae) and the biogeography of transitions between marine and freshwater habitats","volume":"2011","author":"NR Lovejoy","year":"2011","journal-title":"Copeia"},{"issue":"5","key":"pone.0335898.ref014","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1023\/A:1009216030316","article-title":"Chromosomal localization of 5S rDNA genes in Leporinus fish (Anostomidae, Characiformes)","volume":"7","author":"C Martins","year":"1999","journal-title":"Chromosome Res"},{"issue":"1","key":"pone.0335898.ref015","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1006\/mpev.1996.0382","article-title":"Phylogenetic relationships among tick subfamilies (Ixodida: Ixodidae: Argasidae) based on the 18S nuclear rDNA gene","volume":"7","author":"4th Black WC","year":"1997","journal-title":"Mol Phylogenet Evol"},{"issue":"6","key":"pone.0335898.ref016","doi-asserted-by":"crossref","DOI":"10.1101\/pdb.top093336","article-title":"Isolation and Quantification of DNA","volume":"2018","author":"MR Green","year":"2018","journal-title":"Cold Spring Harb Protoc"},{"key":"pone.0335898.ref017","first-page":"0","article-title":"Extra\u00e7\u00e3o de DNA para aplica\u00e7\u00e3o em rea\u00e7\u00e3o em cadeia da polimerase (PCR).","volume-title":"Biologia molecular aplicada \u00e0 produ\u00e7\u00e3o animal","author":"LCA. Regitano","year":"2001"},{"issue":"5","key":"pone.0335898.ref018","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1111\/j.1399-0039.1992.tb01940.x","article-title":"HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation","volume":"39","author":"O Olerup","year":"1992","journal-title":"Tissue Antigens"},{"issue":"10","key":"pone.0335898.ref019","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1016\/j.foodcont.2006.07.016","article-title":"Comparison of DNA extraction methods from muscle of canned tuna for species identification","volume":"18","author":"MJ Chapela","year":"2007","journal-title":"Food Control"},{"key":"pone.0335898.ref020","author":"MCS Oliveira","year":"2007"},{"key":"pone.0335898.ref021","author":"FR Ara\u00fajo","year":"2009"},{"issue":"2","key":"pone.0335898.ref022","doi-asserted-by":"crossref","first-page":"248","DOI":"10.5858\/2008-132-248-MPIAPP","article-title":"Molecular pathology in anatomic pathology practice: a review of basic principles","volume":"132","author":"JL Hunt","year":"2008","journal-title":"Arch Pathol Lab Med"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0335898","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T18:24:28Z","timestamp":1762539868000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0335898"}},"subtitle":[],"editor":[{"given":"Windsor E.","family":"Aguirre","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2025,11,7]]},"references-count":22,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11,7]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0335898","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,7]]}}}