A DNA 'barcode blitz': rapid digitization and sequencing of a natural history collection

PLoS One. 2013 Jul 10;8(7):e68535. doi: 10.1371/journal.pone.0068535. Print 2013.

Abstract

DNA barcoding protocols require the linkage of each sequence record to a voucher specimen that has, whenever possible, been authoritatively identified. Natural history collections would seem an ideal resource for barcode library construction, but they have never seen large-scale analysis because of concerns linked to DNA degradation. The present study examines the strength of this barrier, carrying out a comprehensive analysis of moth and butterfly (Lepidoptera) species in the Australian National Insect Collection. Protocols were developed that enabled tissue samples, specimen data, and images to be assembled rapidly. Using these methods, a five-person team processed 41,650 specimens representing 12,699 species in 14 weeks. Subsequent molecular analysis took about six months, reflecting the need for multiple rounds of PCR as sequence recovery was impacted by age, body size, and collection protocols. Despite these variables and the fact that specimens averaged 30.4 years old, barcode records were obtained from 86% of the species. In fact, one or more barcode compliant sequences (>487 bp) were recovered from virtually all species represented by five or more individuals, even when the youngest was 50 years old. By assembling specimen images, distributional data, and DNA barcode sequences on a web-accessible informatics platform, this study has greatly advanced accessibility to information on thousands of species. Moreover, much of the specimen data became publically accessible within days of its acquisition, while most sequence results saw release within three months. As such, this study reveals the speed with which DNA barcode workflows can mobilize biodiversity data, often providing the first web-accessible information for a species. These results further suggest that existing collections can enable the rapid development of a comprehensive DNA barcode library for the most diverse compartment of terrestrial biodiversity - insects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Australia
  • Biological Specimen Banks*
  • DNA Barcoding, Taxonomic / methods*
  • Feasibility Studies
  • Information Storage and Retrieval / methods
  • Insecta / classification*
  • Insecta / genetics
  • Libraries, Digital*
  • Natural History / methods*
  • Quality Control
  • Sequence Analysis, DNA / methods*
  • Sequence Analysis, DNA / standards
  • Specimen Handling / methods
  • Time Factors

Grants and funding

The authors thank NSERC and CSIRO for the support that allowed the processing of specimens at ANIC. Sequence analysis was enabled by funding from the government of Canada through Genome Canada and the Ontario Genomics Institute in support of the International Barcode of Life project. This work was heavily dependent upon BOLD; the authors thank the Ontario Ministry of Research and Innovation for funding its development. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.