In RNA Mapping- Methods and Protocols expert researchers in the field detail many of the methods which are now commonly used to study RNA. These include protocols for the consequence of the emerging interest in the characterization of cellular RNAs urged by their potential use as diagnostic biomarkers or therapeutic targets. In particular, the biological relevance of microRNAs in human physiology and disease development is highlighted in the 16 chapters focused on methods for their physical and functional mapping. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, RNA Mapping- Methods and Protocols provides instruction and inspiration for scientists who are facing the challenges of the discovery and/or functional characterization of RNA molecules for a wide variety of applications ranging from novel biomedical diagnostics to therapeutics and biomaterials.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Full-Length Characterization Of Transcribed Genomic Regions.- Rapid Mapping Of RNA 3´and 5´Ends.- Single Nucleotide Mapping Of RNA 5´and 3´Ends.- RNA Secondary Structure Analysis.- Tertiary Structure Mapping Of Pri-Mirna17~92.- In Situ Hybridization Detection Of Micrornas Using LNA Oligonucleotides.- Quantification Of Micrornas By A Simple And Specific Qpcr Method.- RNA Isolation For Small RNA Next Generation Sequencing From Acellular Biofluids.- Sequencing Small RNA: Introduction and Data Analysis Fundamentals.- Measuring Expression Levels Of Small Regulatory RNA Molecules From Formalin-Fixed, Paraffin-Embedded Samples and Body Fluids.- Microrna Profiling In Plasma Or Serum Using Quantitative RT-Qpcr.- Microrna Profiling Of Exosomes Isolated From Biofluids And Conditioned Media.- Isolation Of Urinary Exosomes For RNA Biomarker Discovery Using A Simple, Fast, And Highly Scalable Method.- Identification Of Actively Translated Mrnas.- Mapping Of Internal Ribosome Entry Sites (IRES).- Mapping Of Protein Binding RNA Elements.- Purification Of RNA Binding Proteins .- De Novo Approach To Classify Protein-Coding And Non-Coding Transcripts By Utilizing Sequence Intrinsic Composition.- Computational Methods To Predict Long Non-Coding RNA Functions Based On Co-Expression Network.- Microrna Biogenesis - Dicing Assay.- Faster Experimental Validation Of Microrna Targets Using Cold Fusion Cloning And A Dual Firefly-Renilla Luciferase Reporter Assay.- Experimental Validation Of Predicted Mammalian Micrornas Of Mirtron Origin.- A Guide For Microrna Target Prediction And Analysis Using Web-Based Applications.- Tapping Microrna Regulation Networks Through Integrated Analysis Of Microrna-Mrna High-Throughput Profiles.- Mirwalk Database For Microrna-Target Interactions .- A Schematic Workflow For Collecting The Interaction Information Between Copy Number Variants And Micrornas Using Existing Resources.- SYBR Green And Taqman PCR Arrays: Expression Profile Of Genes RelevantTo A Pathway Or Disease State.- Comprehensive Meta-Analysis Of Microrna Expression Using A Robust Rank Aggregation Approach.