This updated edition explores different spectroscopic and biophysical methods used to study the structure and folding of RNA and to follow their interactions with proteins. The book expands into protocols for dynamic and single-molecule analyses, addressing the new challenges that have emerged since the discovery of novel RNAs and their interaction networks. 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, RNA Spectroscopy: Methods and Protocols, Second Edition is a valuable resource for researchers interested in learning more about this vital and growing field of study.Chapter 14 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.