{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T02:51:39Z","timestamp":1791427899267,"version":"4.3.3"},"reference-count":35,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:00:00Z","timestamp":1788480000000},"content-version":"vor","delay-in-days":3,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100020275","name":"Vilnius University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100020275","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Ecology and Evolution"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    No systematic comparison of DNA extraction strategies exists for minute Vertiginidae (shell height &lt;\u20093\u2009mm), a group posing a dual analytical challenge: extremely low tissue input and co\u2010purified PCR\u2010inhibitory mucus. For legally protected species, an additional requirement to preserve the shell voucher further constrains available protocols. Using\n                    <jats:italic>Vertigo antivertigo<\/jats:italic>\n                    as the model species, we compared six approaches applied to specimens preserved in 96% ethanol (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u200910 per method): two HotSHOT alkaline\u2010lysis protocols (destructive and non\u2010destructive shell\u2010preserving variants), a modified CTAB protocol supplemented with PVP\u201040 and DTT, and three commercial silica\u2010column kits (GeneJET Genomic, DNeasy Blood &amp; Tissue, QIAamp DNA Micro). DNA yields were quantified by QuantiFluor fluorometry, and PCR performance was subsequently assessed across four loci (COI barcode, COI mini\u2010barcode, ITS1, ITS2). DNeasy Blood &amp; Tissue produced the highest fluorometric concentrations; QIAamp DNA Micro and CTAB\u2009+\u2009PVP\u201040 gave intermediate values. The shell\u2010preserving HotSHOT variant yielded lower concentrations but improved A260\/230 ratios. BSA and trehalose supplementation increased PCR success in inhibition\u2010prone HotSHOT extracts from 70% to 100%. ITS1 Sanger sequencing of three\n                    <jats:italic>Vertigo<\/jats:italic>\n                    species listed in Annex II of the EU Habitats Directive, all extracted with the shell\u2010preserving protocol, confirmed species\u2010level identification (99.8%\u2013100% BLASTn identity; mean Phred\n                    <jats:italic>Q<\/jats:italic>\n                    \u2009&gt;\u200951). The shell\u2010preserving non\u2010destructive HotSHOT protocol yields sequenceable DNA from protected Vertiginidae while retaining the morphological voucher, making it the preferred option for conservation\u2010genetic monitoring. The practical decision framework documented here\u2014integrating voucher preservation, amplification robustness and per\u2010sample cost\u2014has broad applicability to other minute terrestrial gastropods processed in large\u2010scale biodiversity surveys.\n                  <\/jats:p>","DOI":"10.1002\/ece3.74295","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T09:10:01Z","timestamp":1788513001000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>DNA<\/scp>\n                    Extraction Optimisation for Minute Land Snails of\n                    <i>Vertigo<\/i>\n                    M\u00fcller, 1773 (Gastropoda: Vertiginidae): A Comparative Evaluation of Six Methods, Including a Non\u2010Destructive Shell\u2010Preserving Protocol"],"prefix":"10.1002","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-1724-5624","authenticated-orcid":false,"given":"Benediktas","family":"Jukonis","sequence":"first","affiliation":[{"name":"Department of Zoology, Institute of Biosciences, Life Science Center Vilnius University  Vilnius Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0697-4567","authenticated-orcid":false,"given":"Jekaterina","family":"Havelka","sequence":"additional","affiliation":[{"name":"Department of Zoology, Institute of Biosciences, Life Science Center Vilnius University  Vilnius Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4229-1219","authenticated-orcid":false,"given":"Grita","family":"Skujien\u0117","sequence":"additional","affiliation":[{"name":"Department of Zoology, Institute of Biosciences, Life Science Center Vilnius University  Vilnius Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,4]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2020.0162"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10592\u2010004\u20107823\u20109"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.5962\/p.408170"},{"key":"e_1_2_11_5_1","doi-asserted-by":"crossref","unstructured":"Chakraborty S. A.Saha andA.Neelavar Ananthram.2020. \u201cComparison of DNA Extraction Methods for Non\u2010Marine Molluscs: Is Modified CTAB DNA Extraction Method More Efficient Than DNA Extraction Kits?3Biotech.\u201d10:69.https:\/\/doi.org\/10.1007\/s13205\u2010020\u20102051\u20107.","DOI":"10.1007\/s13205-020-2051-7"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/biomet\/26.4.404"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.6620\/ZS.2024.63\u201019"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1755\u20100998.2009.02559.x"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fevo.2020.00037"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1080\/15257770.2025.2486368"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.9420"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.0014-3820.2002.tb00191.x"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/1755\u20100998.12138"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.4003\/006.038.0209"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1098\/rspb.2002.2218"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102699.2005.01359.x"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/geb.13820"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13104\u2010016\u20102147\u20107"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.3.1102\u20101106.1996"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1186\/1742\u20109994\u201010\u201034"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12686\u2010023\u201001325\u20103"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/biosci\/biaa164"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.3389\/fevo.2024.1329581"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.4002\/040.062.0104"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms12118086"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02772108"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ympev.2022.107561"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.3732\/apps.1200236"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/aps3.11517"},{"key":"e_1_2_11_30_1","first-page":"12","article-title":"A Comparison of Different DNA Extraction Methods for Slugs (Mollusca: Pulmonata)","volume":"1","author":"Skujien\u0117 G.","year":"2003","journal-title":"Ekologija"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2004.031336"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.2144\/00291bm09"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1093\/mollus\/eyw011"},{"key":"e_1_2_11_34_1","first-page":"315","volume-title":"PCR Protocols: A Guide to Methods and Applications","author":"White T. 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