Analysis of rna folding and ligand binding by conventional and high-throughput calorimetry

Joshua E. Sokoloski, Philip C. Bevilacqua

Research output: Chapter in Book/Report/Conference proceedingChapter

7 Scopus citations

Abstract

Noncoding RNAs serve myriad functions in the cell, but their biophysical properties are not well understood. Calorimetry offers direct and label-free means for characterizing the ligand-binding and thermostability properties of these RNA. We apply two main types of calorimetry-isothermal titration calorimetry (ITC) and differential scanning calorimetry (DSC)-to the characterization of these functional RNA molecules. ITC can describe ligand binding in terms of stoichiometry, affinity, and heat (enthalpy), while DSC can provide RNA stability in terms of heat capacity, melting temperature, and folding enthalpy. Here, we offer detailed experimental protocols for studying such RNA systems with commercially available conventional and high-throughput ITC and DSC instruments.

Original languageEnglish (US)
Title of host publicationBacterial Regulatory RNA
Subtitle of host publicationMethods and Protocols
EditorsKenneth C. Keiler
Pages145-174
Number of pages30
DOIs
StatePublished - Jul 27 2012

Publication series

NameMethods in Molecular Biology
Volume905
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Sokoloski, J. E., & Bevilacqua, P. C. (2012). Analysis of rna folding and ligand binding by conventional and high-throughput calorimetry. In K. C. Keiler (Ed.), Bacterial Regulatory RNA: Methods and Protocols (pp. 145-174). (Methods in Molecular Biology; Vol. 905). https://doi.org/10.1007/978-1-61779-949-5_10