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      DNA Extraction from Dry Museum Beetles without Conferring External Morphological Damage

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          Abstract

          Background

          A large number of dry-preserved insect specimens exist in collections around the world that might be useful for genetic analyses. However, until now, the recovery of nucleic acids from such specimens has involved at least the partial destruction of the specimen. This is clearly undesirable when dealing with rare species or otherwise important specimens, such as type specimens.

          Methodology

          We describe a method for the extraction of PCR-amplifiable mitochondrial and nuclear DNA from dry insects without causing external morphological damage. Using PCR to amplify ≈220 bp of the mitochondrial gene cytochrome c oxidase I, and 250–345 bp fragments of the multi-copy, nuclear 28s ribosomal DNA gene, we demonstrate the efficacy of this method on beetles collected up to 50 years ago.

          Conclusions

          This method offers a means of obtaining useful genetic information from rare insects without conferring external morphological damage.

          Related collections

          Most cited references13

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          Molecular Cloning : A Laboratory Manual

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            The promise of a DNA taxonomy.

            Not only is the number of described species a very small proportion of the estimated extant number of taxa, but it also appears that all concepts of the extent and boundaries of 'species' fail in many cases. Using conserved molecular sequences it is possible to define and diagnose molecular operational taxonomic units (MOTU) that have a similar extent to traditional 'species'. Use of a MOTU system not only allows the rapid and effective identification of most taxa, including those not encountered before, but also allows investigation of the evolution of patterns of diversity. A MOTU approach is not without problems, particularly in the area of deciding what level of molecular difference defines a biologically relevant taxon, but has many benefits. Molecular data are extremely well suited to re-analysis and meta-analysis, and data from multiple independent studies can be readily collated and investigated by using new parameters and assumptions. Previous molecular taxonomic efforts have focused narrowly. Advances in high-throughput sequencing methodologies, however, place the idea of a universal, multi-locus molecular barcoding system in the realm of the possible.
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              Nondestructive DNA extraction method for mitochondrial DNA analyses of museum specimens.

              Museum specimens have provided the material for a large proportion of ancient DNA studies conducted during the last 20 years. However, a major drawback of the genetic analyses is that the specimens investigated are usually damaged, as parts of skin, bone, or a tooth have to be removed for DNA extraction. To get around these limitations, we have developed a nondestructive extraction method for bone, tooth, and skin samples. We found that it is possible to amplify mitochondrial DNA (mtDNA) sequences up to at least 414 bp long from samples up to 164 years old. Using this method, almost 90% (35 of 40) of the investigated samples yielded amplifiable mtDNA. Moreover, we found that repeated extractions of the same samples allowed amplifications of the expected length for all samples at least three times and for some samples up to at least five times. Thus this method opens up the possibility to repeatedly use museum collections for mtDNA analyses without damaging the specimens and thus without reducing the value of irreplaceable collections for morphological analyses.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS ONE
                plos
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2007
                7 March 2007
                : 2
                : 3
                : e272
                Affiliations
                [1 ]Department of Ecology and Evolutionary Biology, The University of Arizona, Tucson, Arizona, United States of America
                [2 ]Ancient DNA and Evolution, The Niels Bohr Institute, University of Copenhagen, Copenhagen, Denmark
                [3 ]Department of Entomology, California Academy of Sciences, San Francisco, California, United States of America
                [4 ]Research Laboratory, Institute of Forensic Medicine, The University of Copenhagen, Copenhagen, Denmark
                Max Planck Institute for Evolutionary Anthropology, Germany
                Author notes
                * To whom correspondence should be addressed. E-mail: mtpgilbert@ 123456gmail.com

                Conceived and designed the experiments: MG WM. Performed the experiments: MW MG WM LM. Analyzed the data: MW MG WM LM. Wrote the paper: MW MG WM LM.

                Article
                06-PONE-RA-00312R1
                10.1371/journal.pone.0000272
                1803022
                17342206
                819ff048-80a7-49d6-a022-246f1609667c
                This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
                History
                : 3 November 2006
                : 15 February 2007
                Page count
                Pages: 4
                Categories
                Research Article
                Evolutionary Biology
                Genetics and Genomics/Population Genetics

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