Multiplex PCR Detection of the GT73, MS8xRF3, and T45 Varieties of GM Canola

  • Kim, Jae-Hwan (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Kim, Tae-Woon (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University) ;
  • Lee, Woo-Young (Team of Novel Foods, Korea Food and Drug Administration) ;
  • Park, Sun-Hee (Team of Novel Foods, Korea Food and Drug Administration) ;
  • Kim, Hae-Yeong (Institute of Life Sciences and Resources and Graduate School of Biotechnology, Kyung Hee University)
  • Published : 2007.02.28

Abstract

A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect three varieties of genetically modified (GM) canola. The construct-specific primers were used to distinguish the following three varieties of GM canola; GT73, MS8xRF3, and T45, using multiplex PCR. The FatA (fatty acyl-ACP thioesterase) gene was used as an endogenous canola reference gene in the PCR detection. The primer pair Canendo-FIR containing a 105 bp amplicon was used to amplify the FatA gene and no amplified product was observed in any of the 15 different plants used as templates. The GT73-KHUF1/R1 primer recognized the 3'-flanking region of GT73, resulting in an amplicon of 125 bp. The Barstar-F1/MS8xRF3-R primer recognized the junction region of bars tar and the NOS terminator introduced into MS8xRF3, resulting in a 162 bp amplicon, and the T45-F2/R2 primer recognized the junction region of PAT and the 35S terminator introduced into T45, resulting in an amplicon of 186 bp. This multiplex PCR allowed for the detection of construct-specific targets in a genomic DNA mixture of up to 1% GM canola containing GT73, MS8xRF3, and T45.

Keywords

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