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DNA barcodes from century-old type specimens using next-generation sequencing

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dc.contributor.author Prosser, Sean W. J. en
dc.contributor.author deWaard, Jeremy R. en
dc.contributor.author Miller, Scott E. en
dc.contributor.author Hebert, Paul D. N. en
dc.date.accessioned 2015-10-19T13:58:20Z
dc.date.available 2015-10-19T13:58:20Z
dc.date.issued 2016
dc.identifier.citation Prosser, Sean W. J., deWaard, Jeremy R., Miller, Scott E., and Hebert, Paul D. N. 2016. "<a href="https://repository.si.edu/handle/10088/27422">DNA barcodes from century-old type specimens using next-generation sequencing</a>." <em>Molecular Ecology Resources</em>. 16 (2):487&ndash;497. <a href="https://doi.org/10.1111/1755-0998.12474">https://doi.org/10.1111/1755-0998.12474</a> en
dc.identifier.issn 1755-098X
dc.identifier.uri http://hdl.handle.net/10088/27422
dc.description.abstract Type specimens have high scientific importance because they provide the only certain connection between the application of a Linnean name and a physical specimen. Many other individuals may have been identified as a particular species, but their linkage to the taxon concept is inferential. Because type specimens are often more than a century old and have experienced conditions unfavorable for DNA preservation, success in sequence recovery has been uncertain. The present study addresses this challenge by employing next generation sequencing (NGS) to recover sequences for the barcode region of the cytochrome c oxidase 1 gene from small amounts of template DNA. DNA quality was first screened in more than 1800 century-old type specimens of Lepidoptera by attempting to recover 164bp and 94bp reads via Sanger sequencing. This analysis permitted the assignment of each specimen to one of three DNA quality categories - high (164bp sequence), medium (94bp sequence), or low (no sequence). Ten specimens from each category were subsequently analyzed via a PCR-based NGS protocol requiring very little template DNA. It recovered sequence information from all specimens with average read lengths ranging from 458bp to 610bp for the three DNA categories. By sequencing ten specimens in each NGS run, costs were similar to Sanger analysis. Future increases in the number of specimens processed in each run promise substantial reductions in cost, making it possible to anticipate a future where barcode sequences are available from most type specimens. This article is protected by copyright. All rights reserved. en
dc.relation.ispartof Molecular Ecology Resources en
dc.title DNA barcodes from century-old type specimens using next-generation sequencing en
dc.type Journal Article en
dc.identifier.srbnumber 137491
dc.identifier.doi 10.1111/1755-0998.12474
rft.jtitle Molecular Ecology Resources
rft.volume 16
rft.issue 2
rft.spage 487
rft.epage 497
dc.description.SIUnit NH-Entomology en
dc.description.SIUnit NMNH en
dc.description.SIUnit Peer-reviewed en
dc.citation.spage 487
dc.citation.epage 497


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