Molecular ecology resourcesMol Ecol ResourSuccessful application of ancient DNA extraction and library construction protocols to museum wet collection specimens.229923152299-231510.1111/1755-0998.13433Millions of scientific specimens are housed in museum collections, a large part of which are fluid preserved. The use of formaldehyde as fixative and subsequent storage in ethanol is especially common in ichthyology and herpetology. This type of preservation damages DNA and reduces the chance of successful retrieval of genetic data. We applied ancient DNA extraction and single stranded library construction protocols to a variety of vertebrate samples obtained from wet collections and of different ages. Our results show that almost all samples tested yielded endogenous DNA. Archival DNA extraction was successful across different tissue types as well as using small amounts of tissue. Conversion of archival DNA fragments into single-stranded libraries resulted in usable data even for samples with initially undetectable DNA amounts. Subsequent target capture approaches for mitochondrial DNA using homemade baits on a subset of 30 samples resulted in almost complete mitochondrial genome sequences in several instances. Thus, application of ancient DNA methodology makes wet collection specimens, including type material as well as rare, old or extinct species, accessible for genetic and genomic analyses. Our results, accompanied by detailed step-by-step protocols, are a large step forward to open the DNA archive of museum wet collections for scientific studies.© 2021 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.StraubeNicolasN0000-0001-7047-1084University Museum of Bergen, Bergen, Norway.SNSB Bavarian State Collection of Zoology, München, Germany.LyraMariana LML0000-0002-7863-4965Departamento de Biodiversidade, Instituto de Biociências and Centro de Aquicultura (CAUNESP), Laboratório de Herpetologia, Universidade Estadual Paulista - UNESP, Rio Claro, SP, Brazil.Zoological Institute, Braunschweig University of Technology, Braunschweig, Germany.PaijmansJohanna L AJLA0000-0002-1938-7052Department of Mathematics and Natural Sciences, Evolutionary Adaptive Genomics, Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.PreickMichaelaMDepartment of Mathematics and Natural Sciences, Evolutionary Adaptive Genomics, Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.BaslerNikolasNDepartment of Mathematics and Natural Sciences, Evolutionary Adaptive Genomics, Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.PennerJohannesJMuseum für Naturkunde- Leibniz Institute for Evolution and Biodiversity Science, Berlin, Germany.Chair of Wildlife Ecology and Management, Albert Ludwigs University Freiburg, Freiburg, Germany.RödelMark-OliverMOMuseum für Naturkunde- Leibniz Institute for Evolution and Biodiversity Science, Berlin, Germany.WestburyMichael VMV0000-0003-0478-3930Section for Evolutionary Genomics, The GLOBE Institute, University of Copenhagen, Copenhagen, Denmark.HaddadCélio F BCFBDepartamento de Biodiversidade, Instituto de Biociências and Centro de Aquicultura (CAUNESP), Laboratório de Herpetologia, Universidade Estadual Paulista - UNESP, Rio Claro, SP, Brazil.BarlowAxelADepartment of Mathematics and Natural Sciences, Evolutionary Adaptive Genomics, Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.HofreiterMichaelMDepartment of Mathematics and Natural Sciences, Evolutionary Adaptive Genomics, Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.eng#2013/50741-7Fundação de Amparo à Pesquisa do Estado de São Paulo#2017/2616-8Fundação de Amparo à Pesquisa do Estado de São Paulo#2018/15425-0Fundação de Amparo à Pesquisa do Estado de São Paulo351649567Deutsche Forschungsgemeinschaft431589/2016-0Conselho Nacional de Desenvolvimento Científico e Tecnológico306623/2018-8Conselho Nacional de Desenvolvimento Científico e TecnológicoJournal Article20210616EnglandMol Ecol Resour1014656041755-098X0DNA, Ancient0DNA, MitochondrialIMDNA, AncientDNA, MitochondrialgeneticsMuseumsSequence Analysis, DNASpecimen Handlingancient DNAarchival DNAbiological collectionformalinsingle-stranded DNA librarytarget capture20215920211720215142021527602021918602021526721ppublish3403673210.1111/1755-0998.13433REFERENCES

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