Transgenic Maize: Methods and Protocols

Part I: Introduction1. Design of Gene Constructs for Transgenic MaizeDong LiuPart II: Transformation Methods2. Transient Expression of GFP in Immature Seed TissuesColin T. Shepherd3. Biolistic Gun Mediated Maize Genetic TransformationKan Wang and Bronwyn Frame4. Agrobacterium-Mediated Maize Transformation: Immature Embryos Versus CallusVladimir Sidorov and David Duncan5. Whiskers-Mediated Maize TransformationJoseph F. Petolino and Nicole L. ArnoldPart III: Transgenic Maize in Research6. Methods for Generation and Analysis of Fluorescent Protein Tagged Maize LinesAmitabh Mohanty, Yan Yang, Anding Luo, Anne Sylvester, and David Jackson7. Use of Transgene-Induced RNAi to Regulate Endogenous Gene ExpressionKaren M. McGinnis8. Plasmid Rescue: Recovery of Flanking Genomic Sequences from Transgenic Transposon Insertion SitesGuo-Ling Nan and Virginia WalbotPart IV: Analysis of Transgenic Plants9. Non-Radioactive Genomic DNA Blots for Detection of Low Abundant Sequences in Transgenic MaizeGuo-Ling Nan and Virginia Walbot10. Tissue-Print Immuno-Detection of Transgene Products in Endosperm for High-Throughput Screening of SeedsM. Paul Scott11. Determination of Transgene Copy Number by Real-Time Quantitative-PCRColin T. Shepherd, Adrienne N. Moran Lauter, and M. Paul ScottPart V: Breeding with Transgenes12. Herbicide Resistance Screening AssayJoan M. Peterson13. Characterizing Transgene InheritanceEarl H. Bicar14. Backcross BreedingKarla E. Vogel Appendix A: Sample APHIS Notification of Release of Regulated MaterialAppendix B: Sample SOP for Field Release