Humana Transgenic Plants: Methods and Protocols: 286 (Methods in Molecular Biology)

A collection of readily reproducible techniques for the genetic transformation and regeneration of plants. The authors cover the most commonly used transformation systems for the regeneration of whole transgenic plants. Among the techniques discussed for the detection of transgenes are quantitative real-time PCR, RT-PCR, in situ hybridization, FISH, and TAIL-PCR. Risk assessment methods offer the opportunity to study Agrobacterium persistence in plant tissues and to investigate the possibility of transgene dispersal. The protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. From the Back Cover Transgenic plants serve as essential tools for research in plant biology and are the basis of modern agricultural biotechnology. In Transgenic Plants: Methods and Protocols, leading scientists describe readily reproducible techniques they have perfected for the genetic transformation and regeneration of plants. The authors cover the most commonly used transformation systems, including Agrobacterium tumefaciens, Agrobacterium rhizogenes, particle bombardment, electroporation, floral-dip, and chloroplast transformation, as well as the regeneration of whole transgenic plants by organogenesis and somatic embryogenesis from different explant cells/tissues and from such diverse plant species as tomato, cassava, conifers, and citrus. Also described are the use of selectable and reporter markers, positive selection (for maize transformation), and the controlled excision and removal of marker genes from both nuclei and plastids once transgenic shoots have been efficiently generated. Additional techniques employ quantitative real-time PCR, reverse transcription PCR, fluorescence in situ hybridization, and thermal asymmetric interlaced PCR. Risk assessment methods offer the opportunity to study Agrobacterium persistence in plant tissues and to investigate the possibility of transgene dispersal. The protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and easy to use, Transgenic Plants: Methods and Protocols offers both novice and experienced researchers a cutting-edge laboratory manual covering all aspects of plant genetic transformation and transgenic crops.