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Applied Entomology and Zoology

, Volume 50, Issue 3, pp 419–425 | Cite as

A rapid method of non-destructive DNA extraction from individual springtails (Collembola)

  • Hiroaki Aoyama
  • Seikoh SaitohEmail author
  • Saori Fujii
  • Hideki Nagahama
  • Naoya Shinzato
  • Nobuhiro Kaneko
  • Taizo Nakamori
Technical Note

Abstract

In this study, we describe an easy and rapid method for non-destructive DNA extraction from a single Collembola individual, without dissection, lysis of the specimen, or purification of extracted DNA. We demonstrate that, after a single specimen has been heat-treated in Tris-EDTA (TE) buffer using a standard thermocycler, the solution can be used for PCR amplification of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene region, typically used for DNA barcoding. With this method, the morphological features of Collembola commonly used for species identification are well preserved. This DNA extraction method is preferable for DNA barcoding where the sequencing and preservation of a large number of small and fragile specimens are required.

Keywords

Collembola DNA barcoding Non-destructive extraction Single-individual PCR Voucher 

Notes

Acknowledgments

We thank Kamigamo Experimental Station, Field Science Education and Research Center (FSERC), Kyoto University, Japan, for allowing access to the study site. We are grateful to Dr. K. Ichisawa at the Tottori Prefectural Museum, Tottori, for depositing our specimens in the museum. We thank two anonymous reviewers for their careful reading of the manuscript and constructive comments. This study was partly supported by MEXT/JSPS KAKENHI grant no. 25281029.

Supplementary material

13355_2015_340_MOESM1_ESM.docx (481 kb)
Supplementary material 1 (DOCX 482 kb)

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Copyright information

© The Japanese Society of Applied Entomology and Zoology 2015

Authors and Affiliations

  • Hiroaki Aoyama
    • 1
  • Seikoh Saitoh
    • 1
    Email author
  • Saori Fujii
    • 2
  • Hideki Nagahama
    • 1
  • Naoya Shinzato
    • 1
  • Nobuhiro Kaneko
    • 2
  • Taizo Nakamori
    • 2
  1. 1.Tropical Biosphere Research CenterUniversity of the RyukyusNishiharaJapan
  2. 2.Department of Environment and Natural Sciences, Graduate School of Environment and Information SciencesYokohama National UniversityYokohamaJapan

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