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Applied Entomology and Zoology

, Volume 47, Issue 3, pp 291–294 | Cite as

Simple and quick methods for nematode DNA preparation

  • Ryusei Tanaka
  • Taisei Kikuchi
  • Takuya Aikawa
  • Natsumi KanzakiEmail author
Technical Note

Abstract

A DNA extraction kit, ISOHAIR® (Nippon Gene), which was originally developed for preparing DNA from hair and nail samples, was used to prepare nematode DNA for PCR and sequencing analyses. The methods provided here, which involved digesting (resolving) a single nematode individual in a tube containing the mixed enzyme solution, enabled the DNA to be prepared within 20 min. The prepared DNA was suitable for PCR amplification of near-full-length small subunit ribosomal RNA (ca 1.7 kb), of the D2/D3 expansion segments of large subunit RNA (ca. 0.7 kb), and of partial mitochondrial COI (ca. 0.7 kb) genes, followed by sequencing analysis. Furthermore, the prepared material could be preserved in a freezer (−30 °C) for at least 20 months, and more than 300 PCR reactions could be performed from a single individual nematode.

Keywords

DNA ISOHAIR® Nematode PCR Sequencing 

Supplementary material

13355_2012_115_MOESM1_ESM.xlsx (18 kb)
Supplementary material 1 (XLSX 18 kb)

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Copyright information

© The Japanese Society of Applied Entomology and Zoology 2012

Authors and Affiliations

  • Ryusei Tanaka
    • 1
  • Taisei Kikuchi
    • 1
  • Takuya Aikawa
    • 2
  • Natsumi Kanzaki
    • 1
    Email author
  1. 1.Forest Pathology LaboratoryForestry and Forest Products Research Institute (FFPRI)TsukubaJapan
  2. 2.Tohoku Research CenterFFPRIMoriokaJapan

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