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This second edition provides updated and comprehensive methods on miRNA biogenesis and their role in the development and progression of various human diseases. Chapters detail miRNA biogenesis, isolating RNA, extracellular vesicles (EVs), circulating miRNAs, analyzing miRNA and miRDeep-P2, protocols for total RNA isolation from cells, cell-derived products, isolation and characterization of exosomes, serum, plasma specimens, and software tools. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, MicroRNA Profiling: Methods and Protocols, Second Edition aims to provide comprehensive and accessible methods to undergraduate, graduate, and established scientist.
List of contents
miRNA Biogenesis and Regulation of Diseases: An updated Overview.- Exosomal MicroRNA profiling.- MicroRNA expression profiling using Agilent One-Color Microarray.- A simple method for profiling and analysing microRNAs from small volume samples using a qPCR-based platform.- Exosomal microRNAs: comprehensive methods from exosome isolation to miRNA extraction and purity analysis.- Detection of MicroRNAs in Brain Slices Using In Situ Hybridization.- MicroRNA profiling of cell lines and xenografts by quantitative PCR.- Assessment of basic biological functions exerted by miRNAs.- Serum microRNAs Profiling in Age-Related Macular Degeneration.- Exosomal microRNA discovery in Age-Related Macular Degeneration.- MicroRNA profiling using a PCR based method.- Guidelines on designing microRNA sponges: from construction to stable cell line.- Evaluating single-nucleotide variants in microRNAs targeting sites and mature microRNAs in vitro cell culture by luciferase reporter gene assays.- Assessment of cell cytotoxicity in 3D biomaterial scaffolds following miRNA transfection.- Evaluation of miRNA expression in 3D in vitro scaffold-based cancer models.- Bioinformatics Analysis of MiRNA sequencing Data.- Plant microRNA identification and annotation using deep sequencing data.
