This updated book covers a wide range of techniques on RNA extraction, detection, quantification, visualization, and genome-wide profiling, from conventional methods to state-of-the-art high throughput approaches. It includes detailed techniques to examine mRNAs, small non-coding RNAs, protein-associated small RNAs, organelle RNAs, endosymbiont RNAs, and alternatively spliced RNA variants from various organisms. RNA editing and the computational data processing for genome-wide datasets is also discussed. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, RNA Abundance Analysis: Methods and Protocols, Second Edition provides a comprehensive set of techniques and methods on isolating and analyzing mRNAs, small RNAs, and other RNA variants, in order to assist researchers in their gene expression studies.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in Nicotiana benthamiana .- Strand-Specific RNA-Seq Applied to Malaria Samples.- Laser Microdissection of Cells and Isolation of High-Quality RNA after Cryosectioning.- Detection of RNA in Ribonucleoprotein Complexes by Blue Native Northern Blotting.- Quantitative Analysis of Plant miRNA Primary Transcripts.- A Revised Adaptation of the Smart-Seq2 Protocol for Single Nematode RNA-Seq.- Analysis of RBP Regulation and Co-Regulation of mRNA 3' UTR Regions in a Luciferase Reporter System.- Extraction of Small RNAs by Titanium Dioxide Nanofibers.- Identification of MicroRNAs and Natural Antisense Transcript-Originated Endogenous siRNAs from Small-RNA Deep Sequencing Data.- Purification and Analysis of Chloroplast RNAs in Arabidopsis.- In Situ Detection of Mature miRNAs in Plants Using LNA-Modified DNA Probes.- Northern Blotting Technique for Detection and Expression Analysis of mRNAs and Small RNAs.- Isolation of Insect Bacteriocytes as a Platform for Transcriptomic Analyses.- Small RNA Isolation and Library Construction for Expression Profiling of Small RNAs from Neurospora crassa and Fusarium oxysporum and Analysis of Small RNAs in Fusarium oxysporum -Infected Plant Root Tissue.- Studying RNA-Protein Interaction Using Riboproteomics.- Small RNA Extraction and Quantification of Isolated Fungal Cells from Plant Tissue by the Sequential Protoplastation.