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United States Department of Agriculture

Agricultural Research Service

Research Project: BIORATIONAL TECHNOLOGIES FOR MANAGEMENT OF CHRYSOMELID BEETLE PESTS OF AGRICULTURAL CROPS Title: Validating DNA barcodes: A non-destructive extraction protocol enables simultaneous vouchering of DNA and morphological vouchers

Authors
item Rowley, Daniel
item Coddington, Jonathan - U.S. NATIONAL MUSEUM
item Gates, Michael
item Norrbom, Allen
item Ochoa, Ronald
item Vandenberg, Natalia
item Greenstone, Matthew

Submitted to: National Entomological Society of America Annual Meeting
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 27, 2007
Publication Date: September 1, 2007
Citation: Rowley, D.L., Coddington, J.A., Gates, M.W., Norrbom, A.L., Ochoa, R., Vandenberg, N.J., Greenstone, M.H. 2007. Validating DNA barcodes: A non-destructive extraction protocol enables simultaneous vouchering of DNA and morphological vouchers. National Entomological Society of America Annual Meeting. 7:915-924.

Technical Abstract: Morphology-based keys support accurate identification of many taxa. However, identification can be difficult for taxa that are not well studied, very small, members of cryptic species complexes, or represented by immature stages. For such cases, DNA barcodes may provide diagnostic characters. Ecologists and evolutionary biologists deposit museum vouchers to document the species studied in their research. If DNA barcodes are to be used for identification, then both the DNA and the specimen from which it was extracted should be vouchered. We describe a protocol for the non-destructive extraction of DNA from terrestrial arthropods, using as examples members of the orders Acarina, Araneae, Coleoptera, Diptera, and Hymenoptera chosen to represent the ranges in size, overall sclerotization, and delicacy of key morphological characters in the group. We successfully extracted sequenceable DNA from all species after 1 – 4 h of immersion in extraction buffer. The extracted carcasses, processed and imaged using protocols standard for the taxon, were distinguishable from closely related species, and adequate as morphological vouchers. We provide links from the carcasses and DNA vouchers to image (MorphBank) and sequence (GenBank) databases.

Last Modified: 12/21/2014
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