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A novel detection system for the genetically modified canola (Brassica rapa) line RT73.

Authors
  • Akiyama, Hiroshi
  • Makiyama, Daiki
  • Nakamura, Kosuke
  • Sasaki, Nobuhiro
  • Minegishi, Yasutaka
  • Mano, Junichi
  • Kitta, Kazumi
  • Ozeki, Yoshihiro
  • Teshima, Reiko
Type
Published Article
Journal
Analytical Chemistry
Publisher
American Chemical Society
Publication Date
Dec 01, 2010
Volume
82
Issue
23
Pages
9909–9916
Identifiers
DOI: 10.1021/ac102434q
PMID: 21049930
Source
Medline
License
Unknown

Abstract

The herbicide-tolerant genetically modified Roundup Ready canola (Brassica napus) line RT73 has been approved worldwide for use in animal feed and human food. However, RT73 Brassica rapa lines derived from interspecific crosses with RT73 B. napus have not been approved in Japan. Here, we report on a novel system using individual kernel analyses for the qualitative detection of RT73 B. rapa in canola grain samples. We developed a duplex real-time polymerase chain reaction (PCR) method to discriminate B. napus and B. rapa DNA using scatter plots of the end-point analyses; this method was able to discriminate a group comprising B. rapa and Brassica juncea from a group comprising B. napus, Brassica carinata, and Brassica oleracea. We also developed a duplex real-time PCR method for the simultaneous detection of an RT73-specific sequence and an endogenous FatA gene. Additionally, a DNA-extraction method using 96-well silica-membrane plates was developed and optimized for use with individual canola kernels. Our detection system could identify RT73 B. rapa kernels in canola grain samples enabling the accurate and reliable monitoring of RT73 B. rapa contamination in canola, thus playing a role in its governmental regulation in Japan.

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