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This detailed volume examines classical and cutting-edge methods involving double-stranded RNA (dsRNA), specifically regarding isolation, visualization, characterization, production, and application. Many protocols, such as co-immunoprecipitation-based isolation of double-stranded RNA-associated protein complexes, identification of mycoviruses by dsRNA extraction, application of dsRNA for fungi disease management (Sclerotinia sclerotiorum and Botrytis cin), and production of double-stranded RNA in plants by plant viral vectors for gene silencing, can also be easily adapted for identification of viruses from other organisms, control of other pathogens, and fundamental research. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips for troubleshooting and avoiding known pitfalls.
Reliable and practical, Double-Stranded RNA: Methods and Protocols serves as an ideal reference book for students and researchers who work with dsRNA.
List of contents
Isolation of Double-Stranded RNAs by Lithium Chloride Fractionation.- Detection of dsRNA by Acridine Orange Staining.- Isolation of dsRNA from Plants by Cellulose Chromatography.- Rapid Purification of dsRNA Using Micro-Spin Cellulose Column.- Analysis of Virus-Induced Double-Stranded RNA in Living Plant Cells by the dRBFC Assay.- Detection of dsRNA with the Fluorescence In Situ Hybridization (FISH) Assay.- Subcellular Colocalization Assay of Host Factors with Viral Replication Complex in the dsRNA Reporter Nicotiana benthamiana.- Production of Double-Stranded RNA Using the Prokaryotic Promoter-Mediated Bidirectional Transcription.- Inducible Expression of dsRNA in Escherichia coli.- In Vivo Production of dsRNA Using Bacteriophage 6 in Pseudomonas syringae Cit7 Cells.- Bacteria-Based Double-Stranded RNA Production to Develop Cost-Effective RNA Interference Application for Insect Pest Management.- Transiently Induce RNA Silencing in Plants Using a TobaccoNecrosis Virus A (TNV-A)-Based dsRNA Production System.- Co-Immunoprecipitation-Based Isolation of Double-Stranded RNA-Associated Protein Complexes in Nicotiana benthamiana.- Analysis of Plant Virus-Induced Immunity by Using Viral-Derived Double-Stranded RNA in Arabidopsis thaliana.- Identification of Mycoviruses by dsRNA Extraction.- Production of Double-Stranded RNA In Planta by a Potato Mop-Top Virus (PMTV)-Based Vector for Inducing Gene Silencing.- Application of dsRNA for Fungi Disease Management Sclerotinia sclerotiorum and Botrytis cinerea.- Application of dsRNA in the Pine Wood Nematode, Bursaphelenchus xylophilus.
