Successful application of ancient DNA extraction and library construction protocols to museum wet collection specimens

Made available in DSpace on 2022-04-29T08:29:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Deutsche Forschungsgemeinschaft Millions 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. University Museum of Bergen SNSB Bavarian State Collection of Zoology Departamento de Biodiversidade Instituto de Biociências and Centro de Aquicultura (CAUNESP) Laboratório de Herpetologia Universidade Estadual Paulista - UNESP Zoological Institute Braunschweig University of Technology Department of Mathematics and Natural Sciences Evolutionary Adaptive Genomics Institute for Biochemistry and Biology University of Potsdam Museum für Naturkunde– Leibniz Institute for Evolution and Biodiversity Science Chair of Wildlife Ecology and Management Albert Ludwigs University Freiburg Section for Evolutionary Genomics The GLOBE Institute University of Copenhagen Department of Zoology Cambridge University Department of Microbiology Immunology and Transplantation Division of Clinical and Epidemiological Virology Rega Institute for Medical Research School of Science and Technology Nottingham Trent University Departamento de Biodiversidade Instituto de Biociências and Centro de Aquicultura (CAUNESP) Laboratório de Herpetologia Universidade Estadual Paulista - UNESP FAPESP: #2013/50741-7 FAPESP: #2017/2616-8 FAPESP: #2018/15425-0 CNPq: 306623/2018-8 Deutsche Forschungsgemeinschaft: 351649567 CNPq: 431589/2016-0