Arabidopsis Protocols, 2nd Edition

This collection of readily reproducible Arabidopsis protocols has been updated to reflect recent advances in plant biology, the completion of the Arabidopsis genome sequence-essential for studying plant function, and the development of whole systems approaches that allow global analysis of gene expression and protein and metabolite dynamics. The authors have included nearly all techniques developed in Arabidopsis, others recently adapted from the traditional work in crop species, and the most recent ones using Arabidopsis as a model system.

Includes supplementary material: sn.pub/extras

Klappentext
Emulating the much-praised first edition of Arabidopsis Protocols, leading scientists have generated an up-to-date work that reflects recent advances in plant biology, the completion of the Arabidopsis genome sequence-essential for studying plant function-and the development of whole systems approaches that allow global analysis of gene expression, as well as protein and metabolite dynamics. The authors have included nearly all techniques developed in Arabidopsis, others recently adapted from more traditional work in crop species, and the latest using Arabidopsis as a model system. Highlights include the most recent methods-transcriptomics, proteomics, and metabolomics-and their novel applications (phosphoproteomics, DNA microarray-based genotyping, high throughput metabolite profiling, and single-cell RNA). Traditional protocols from the agricultural sciences and others developed in crop species (grafting and chloroplast transformation) have been adapted to exploit the advantages of the Arabidopsis model. The protocols themselves follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Indispensable and highly practical, Arabidopsis Protocols, Second Edition offers both novice and experienced plant biologists cutting-edge tools to explore new scenarios and gain an understanding of how this complex, multicellular organism works, how it copes with a sessile life style, and how these strategies compare with those developed in other organisms.

Inhalt
Growing Arabidopsis.- Growth of Plants and Preservation of Seeds.- Prevention and Control of Pests and Diseases.- Growth of Other Species Related to Arabidopsis thaliana.- Grafting.- Synchronization, Transformation, and Cryopreservation of Suspension-Cultured Cells.- Genetic Analyses.- Genetic Analysis.- QTL Analysis.- EMS Mutagenesis of Arabidopsis.- Low-Resolution Mapping of Untagged Mutations.- Gene Identification and Cloning by Molecular Marker Mapping.- High-Throughput TILLING for Arabidopsis.- Genotyping and Mapping With High-Density Oligonucleotide Arrays.- Forward Genetic Screening of Insertional Mutants.- PCR-Based Screening for Insertional Mutants.- Cytogenetic Analyses of Arabidopsis.- Using Information From Public Arabidopsis Databases to Aid Research.- Transient and Stable Transformation.- In Planta Agrobacterium-Mediated Transformation by Vacuum Infiltration.- Transient Expression Assay by Agroinfiltration of Leaves.- Functional Analysis of Transcription Factors by Microparticle Bombardment.- Transfection Assays With Protoplasts Containing Integrated Reporter Genes.- Chloroplast Genetic Engineering Via Organogenesis or Somatic Embryogenesis.- ?-Glucuronidase as Reporter Gene.- Use of Fluorescent Proteins as Reporters.- Luciferases as Reporter Genes.- Aequorin as a Reporter Gene.- Applications of Chemical-Inducible Expression Systems in Functional Genomics and Biotechnology.- Transcriptomics.- RNA Extraction.- Target Preparation for DNA Microarray Hybridization.- Statistical Issues in Microarray Data Analysis.- Preparation and Quality Assessment of RNA From Cell-Specific Samples Obtained by Laser Microdissection.- Proteomics.- Protein Isolation and Second-Dimension Electrophoretic Separation.- Isolation of Nuclear Proteins.- Purification and Fractionationof Membranes for Proteomic Analyses.- Protein Microsequencing.- Peptide-Based Phosphoproteomics With Immobilized Metal Ion Chromatography.- Metabolomics.- Metabolite Profiling in Arabidopsis.- Hormone Profiling in Arabidopsis.