Vouchering DNA-Barcoded Specimens: Test of a Nondestructive Extraction Protocol for Terrestrial Arthropods

Authors

Daniel L. Rowley, United States Department of Agriculture, Agricultural Research Service, Insect Biocontrol Laboratory, Bldg. 011 A, Room 214, BARC-West, Beltsville, Maryland 20705, USA
Jonathan A. Coddington, Smithsonian Institution, National Museum of Natural History, Washington, D.C., USA
Michael W. Gates, United States Department of Agriculture, Agricultural Research Service, Systematic Entomology Laboratory, Beltsville, Maryland, USA
Allen L. Norrbom, United States Department of Agriculture, Agricultural Research Service, Systematic Entomology Laboratory, Beltsville, Maryland, USA
Ronald A. Ochoa, United States Department of Agriculture, Agricultural Research Service, Systematic Entomology Laboratory, Beltsville, Maryland, USA
Natalia J. Vandenberg, United States Department of Agriculture, Agricultural Research Service, Systematic Entomology Laboratory, Beltsville, Maryland, USA
Matthew H. Greenstone, United States Department of Agriculture, Agricultural Research Service, Insect Biocontrol Laboratory, Bldg. 011 A, Room 214, BARC-West, Beltsville, Maryland 20705, USA

Date of this Version

2007

Comments

Published in Molecular Ecology Notes (2007) 7 , 915–924.

Abstract

Morphology-based keys support accurate identification of many taxa. However, identification can be difficult for taxa that are either 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 nondestructive 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.