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This second edition provides new and updated chapters detailing methods tackling all aspects of small non-coding RNAs biology. Chapters guide readers through customized dedicated protocols and technologies that will be of valuable help decipher the numerous functions of small non-coding RNAs. Written in the highly successful Methods of 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.
Instructive and practical, Small Non-Coding RNAs: Methods and Protocols, Second Edition aims to be a valuable help and of great interest to a large number of researchers working in the field of RNA or willing to develop RNA studies.
List of contents
Regulatory Non-coding RNAs: An Overview.- RNA Precipitation.- Quantitative and Qualitative Assessment of Small RNA Preparations.- Non-coding RNA Silencing in Mammalian Cells by Antisense LNA GapmeRs Transfection.- Northern Blot Detection of Tiny RNAs.- Stem-loop qRT-PCR Based Quantification of miRNAs.- Detection and Labeling of Small Non-coding RNAs by Splinted Ligation.- Fluorescence In Situ Hybridization Of Small Non-coding RNAs.- Using native RIP, UV-CLIP or fCLIP to Address Protein-RNA Interactions In Vivo.- Purification of RiboNucleoProtein Particles by MS2-MBP Affinity Chromatography.- Study of genome-wide occupancy of long non-coding RNAs using Chromatin Isolation by RNA Purification (ChIRP).- Gene Reporter Assays to Study miRNA Function.- CRISPR/Cas9 System to Knock-Down microRNA In Vitro and In Vivo.- Microarray Analysis of Whole-Transcriptome RNAs including Non-Coding RNAs.- Isolation, Extraction, and Deep-Sequencing Analysis Of Extracellular RNAs (exRNAs) from Human Plasma.- Analysis of the RNA and protein complexome by Grad-seq.- µIVC-Seq: A Method for Ultrahigh-throughput Development and Functional Characterization of Small RNAs.- RNA Secondary Structure Study by Chemical Probing Methods using DMS and CMCT.- Application of NMR Spectroscopy to Determine the 3D structure of Small Non-coding RNAs.
