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Screening of Genetically Modified Foods by Single-Tube Semi-Nested PCR

YAN Wei1, XU Zhenhui2, LONG Likun1, LI Congcong1, LI Feiwu1,*   

  1. 1. Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, Changchun 130033, China;
    2. College of Life Science and Technology, Harbin Normal University, Harbin 150025, China
  • Online:2015-01-25 Published:2015-01-16
  • Contact: LI Feiwu

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Abstract:

A single-tube semi-nested PCR method for common exogenous regulatory elements of transgenic crops was
established to provide a fast and accurate assay for screening genetically modified foods. Two sets of element-specific nested
PCR primers were designed based on the consensus sequence of the exogenous elements in six different transgenic crops,
respectively. A single-tube semi-nested PCR assay for CaMV35S promoter and NOS terminator was developed by testing
the amplification efficiency of different combinations of primers. This assay has been successfully applied to distinguish
GM crops with CaMV35S promoter or NOS terminator from others with high specificity. Sensitivity tests showed that the
relative limit of detection for CaMV35S promoter and NOS terminator were 0.01% and 0.05%, respectively, significantly
better than those of conventional PCR. Consequently, the single-tube semi-nested PCR assay is convenient, accurate,
sensitive and suitable for screening genetically modified foods.

Key words: genetically modified foods, single-tube semi-nested PCR, screening and detection

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