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This article in TPG

  1. Vol. 5 No. 3, p. 149-163
    unlockOPEN ACCESS
     
    Received: Oct 11, 2012
    Published: December 12, 2012


    * Corresponding author(s): david.k.kovalic@monsanto.com
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doi:10.3835/plantgenome2012.10.0026

The Use of Next Generation Sequencing and Junction Sequence Analysis Bioinformatics to Achieve Molecular Characterization of Crops Improved Through Modern Biotechnology

  1. David Kovalic a,
  2. Carl Garnaata,
  3. Liang Guob,
  4. Yongpan Yana,
  5. Jeanna Groata,
  6. Andre Silvanovicha,
  7. Lyle Ralstona,
  8. Mingya Huanga,
  9. Qing Tiana,
  10. Allen Christiana,
  11. Nordine Cheikha,
  12. Jerry Hjellea,
  13. Stephen Padgettea and
  14. Gary Bannona
  1. a Monsanto Company, 800 N. Lindbergh Blvd., St. Louis, MO 63167
    b Bayer Crop Science, 2 T.W. Alexander Dr., RTP, NC 27709

Abstract

The assessment of genetically modified (GM) crops for regulatory approval currently requires a detailed molecular characterization of the DNA sequence and integrity of the transgene locus. In addition, molecular characterization is a critical component of event selection and advancement during product development. Typically, molecular characterization has relied on Southern blot analysis to establish locus and copy number along with targeted sequencing of polymerase chain reaction products spanning any inserted DNA to complete the characterization process. Here we describe the use of next generation (NexGen) sequencing and junction sequence analysis bioinformatics in a new method for achieving full molecular characterization of a GM event without the need for Southern blot analysis. In this study, we examine a typical GM soybean [Glycine max (L.) Merr.] line and demonstrate that this new method provides molecular characterization equivalent to the current Southern blot-based method. We also examine an event containing in vivo DNA rearrangement of multiple transfer DNA inserts to demonstrate that the new method is effective at identifying complex cases. Next generation sequencing and bioinformatics offers certain advantages over current approaches, most notably the simplicity, efficiency, and consistency of the method, and provides a viable alternative for efficiently and robustly achieving molecular characterization of GM crops.

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Copyright © 2012. Copyright © 2012 Crop Science Society of America