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Analytical and Bioanalytical Chemistry

, Volume 396, Issue 6, pp 2091–2102 | Cite as

Detection and identification of multiple genetically modified events using DNA insert fingerprinting

  • Philippe RaymondEmail author
  • Louis Gendron
  • Moustafa Khalf
  • Sylvianne Paul
  • Kim L. Dibley
  • Somanath Bhat
  • Vicki R. D. Xie
  • Lina Partis
  • Marie-Eve Moreau
  • Cheryl Dollard
  • Marie-José Coté
  • Serge Laberge
  • Kerry R. Emslie
Original Paper

Abstract

Current screening and event-specific polymerase chain reaction (PCR) assays for the detection and identification of genetically modified organisms (GMOs) in samples of unknown composition or for the detection of non-regulated GMOs have limitations, and alternative approaches are required. A transgenic DNA fingerprinting methodology using restriction enzyme digestion, adaptor ligation, and nested PCR was developed where individual GMOs are distinguished by the characteristic fingerprint pattern of the fragments generated. The inter-laboratory reproducibility of the amplified fragment sizes using different capillary electrophoresis platforms was compared, and reproducible patterns were obtained with an average difference in fragment size of 2.4 bp. DNA insert fingerprints for 12 different maize events, including two maize hybrids and one soy event, were generated that reflected the composition of the transgenic DNA constructs. Once produced, the fingerprint profiles were added to a database which can be readily exchanged and shared between laboratories. This approach should facilitate the process of GMO identification and characterization.

Keywords

Genetically modified organism detection GMO Anchored PCR Genome walking Fingerprint 

Notes

Acknowledgments

Work conducted at the National Measurement Institute, Australia was funded by the Office of the Gene Technology Regulator, Australia. CFIA research was funded by the Canadian Regulatory System for Biotechnology (CRSB). We thank Kate Griffiths, Carole Simard, and André Perron for their constructive review of the manuscript.

Supplementary material

216_2009_3295_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 408 kb)

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Copyright information

© Crown copyright in the right of Canada 2009

Authors and Affiliations

  • Philippe Raymond
    • 1
    Email author
  • Louis Gendron
    • 1
  • Moustafa Khalf
    • 1
  • Sylvianne Paul
    • 1
  • Kim L. Dibley
    • 2
  • Somanath Bhat
    • 2
  • Vicki R. D. Xie
    • 2
  • Lina Partis
    • 2
  • Marie-Eve Moreau
    • 1
  • Cheryl Dollard
    • 3
  • Marie-José Coté
    • 3
  • Serge Laberge
    • 4
  • Kerry R. Emslie
    • 2
  1. 1.St-Hyacinthe LaboratoryCanadian Food Inspection Agency (CFIA)St-HyacintheCanada
  2. 2.National Measurement Institute (NMI)PymbleAustralia
  3. 3.Ottawa Laboratory (Fallowfield)Canadian Food Inspection AgencyOttawaCanada
  4. 4.Centre de Recherches de Sainte-FoyAgriculture and Agrifood CanadaSainte-FoyCanada

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