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Molecular Analyses of Transgenic Plants

  • Kurniawan Rudi Trijatmiko
  • Felichi Mae Arines
  • Norman Oliva
  • Inez Hortense Slamet-Loedin
  • Ajay KohliEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1385)

Abstract

One of the major challenges in plant molecular biology is to generate transgenic plants that express transgenes stably over generations. Here, we describe some routine methods to study transgene locus structure and to analyze transgene expression in plants: Southern hybridization using DIG chemiluminescent technology for characterization of transgenic locus, SYBR Green-based real-time RT-PCR to measure transgene transcript level, and protein immunoblot analysis to evaluate accumulation and stability of transgenic protein product in the target tissue.

Keywords

Transgene Transformation Gene expression qRT-PCR Southern blot Western blot Digoxigenin Chemiluminescent SYBR Green 

Notes

Acknowledgements

The authors would like to thank Lina Torrizo, Marina Manzanilla, and Maria Gina Borja for help with rice transformation. Work at the GTL is supported by HarvestPlus.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Kurniawan Rudi Trijatmiko
    • 1
  • Felichi Mae Arines
    • 1
  • Norman Oliva
    • 1
  • Inez Hortense Slamet-Loedin
    • 1
  • Ajay Kohli
    • 2
    Email author
  1. 1.Genetic Transformation Laboratory, Plant Breeding, Genetics and Biotechnology DivisionInternational Rice Research InstituteMetro ManilaPhilippines
  2. 2.Plant Molecular Biology Laboratory, Plant Breeding, Genetics and Biotechnology DivisionInternational Rice Research InstituteMetro ManilaPhilippines

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