{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T03:35:00Z","timestamp":1789702500781,"version":"4.0.1"},"reference-count":48,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T00:00:00Z","timestamp":1748908800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Cell Signalling Technology Inc. Annual Grant","award":["NA"],"award-info":[{"award-number":["NA"]}]},{"name":"Donald Comb Research Scholarship Fund","award":["NA"],"award-info":[{"award-number":["NA"]}]},{"name":"Ocean Genome Legacy Operations Fund","award":["NA"],"award-info":[{"award-number":["NA"]}]},{"name":"Francis Goelet Charitable Lead Trust","award":["NA"],"award-info":[{"award-number":["NA"]}]},{"DOI":"10.13039\/100000153","name":"Division of Biological Infrastructure","doi-asserted-by":"publisher","award":["1722553"],"award-info":[{"award-number":["1722553"]}],"id":[{"id":"10.13039\/100000153","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Cryopreservation is the gold standard for preserving high molecular weight (HMW) DNA (&gt;10\u2009kb) in tissue samples. However, frozen tissues are typically thawed either before or during DNA extraction, which can lead to substantial DNA degradation. In this study, we thawed the previously frozen tissues of 10 marine species (five fishes and five invertebrates) in the preservatives EDTA (250\u2009mM, pH 10) or ethanol (EtOH, 95%) and maintained them in their respective preservatives overnight at 4\u00b0C before DNA extraction. We then compared the recovery of HMW DNA in these extracts to extracts prepared directly from frozen tissues. To evaluate the effect of these treatments on HMW DNA recovery, we determined the percentage of high molecular weight DNA (%HMW) and yield of HMW DNA normalized by tissue weight (nY) in each DNA extract. The average %HMW values for eight of the 10 species and the average nY values for five of the 10 species were significantly higher in extracts from EDTA-treated tissues compared to extracts from untreated frozen tissues. For all 10 species, we observed no significant decreases in average %HMW or nY values in extracts of EDTA-thawed tissues compared to those extracted directly from frozen tissues. In contrast, EtOH treatment did not significantly improve the average %HMW or nY values in extracts from tissues of nine of the 10 species when compared to extracts prepared directly from frozen tissues. Therefore, investigators may consider EDTA treatment as a simple method for improving HMW DNA recovery from frozen tissues.<\/jats:p>","DOI":"10.1371\/journal.pone.0321872","type":"journal-article","created":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T17:36:01Z","timestamp":1748972161000},"page":"e0321872","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Perish the thawed? EDTA reduces DNA degradation during extraction from frozen tissue"],"prefix":"10.1371","volume":"20","author":[{"given":"Ella","family":"Messner","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lev","family":"Becker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-7395-4745","authenticated-orcid":true,"given":"Mia L.","family":"DeSanctis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth A.","family":"Soranno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-0835-7612","authenticated-orcid":true,"given":"Ryan","family":"Pianka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7918-9842","authenticated-orcid":true,"given":"Caileigh","family":"Pierce","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7063-0520","authenticated-orcid":true,"given":"Molly","family":"Johnson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0297-4506","authenticated-orcid":true,"given":"Rosalia Falco","family":"Poulin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8408-7729","authenticated-orcid":true,"given":"Hannah J.","family":"Appiah-Madson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3860-194X","authenticated-orcid":true,"given":"Daniel L.","family":"Distel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,3]]},"reference":[{"issue":"23","key":"pone.0321872.ref001","doi-asserted-by":"crossref","first-page":"5935","DOI":"10.1111\/mec.15909","article-title":"Opportunities and challenges for high-quality biodiversity tissue archives in the age of long-read sequencing","volume":"30","author":"MPK Blom","year":"2021","journal-title":"Mol Ecol"},{"key":"pone.0321872.ref002","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1146\/annurev-genet-071719-020506","article-title":"Museum genomics","volume":"55","author":"D Card","year":"2021","journal-title":"Annu Rev Genet"},{"issue":"1","key":"pone.0321872.ref003","doi-asserted-by":"crossref","first-page":"1246","DOI":"10.1038\/s42003-023-05615-2","article-title":"Generating high-quality plant and fish reference genomes from field-collected specimens by optimizing preservation","volume":"6","author":"JJ Minich","year":"2023","journal-title":"Commun Biol"},{"issue":"5","key":"pone.0321872.ref004","doi-asserted-by":"crossref","first-page":"1690","DOI":"10.1093\/jmammal\/gyz111","article-title":"Curatorial guidelines and standards of the American Society of Mammalogists for collections of genetic resources","volume":"100","author":"CD Phillips","year":"2019","journal-title":"J Mammalogy"},{"key":"pone.0321872.ref005","doi-asserted-by":"crossref","first-page":"77","DOI":"10.14351\/0831-0005-28.1.77","article-title":"Best practices for genetic resources associated with natural history collections: Recommendations for practical implementation","volume":"28","author":"BM Zimkus","year":"2014","journal-title":"Collection Forum"},{"issue":"2","key":"pone.0321872.ref006","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1007\/s12686-011-9545-x","article-title":"Guidelines for collecting and maintaining archives for genetic monitoring","volume":"4","author":"JA Jackson","year":"2011","journal-title":"Conservation Genet Resour"},{"key":"pone.0321872.ref007","unstructured":"Zimkus BM, Ford LS, editors. 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