Recent advancements in calorimetric methods have enhanced the study of RNA folding transitions, enabling high-precision structural analysis. Among these methods, Differential Scanning Calorimetry (DSC) measures the excess heat capacity (Cp) of nucleic acids in solution by detecting differential heat flow between the sample and a reference as the temperature is increased at a constant rate. This technique captures folding or unfolding reactions, with peaks in DSC thermograms indicating transitions and melting points (Tm). Integrating these peaks provides the enthalpy change (ΔH), while the change in heat capacity (ΔCp) is derived from baseline differences. Overall, DSC is a powerful tool for detailed thermodynamic characterization of RNA, offering precise, direct, and comprehensive data without the need for sample modification—allowing for label-free measurements. Furthermore, DSC is versatile, enabling the study of RNA in the presence of various solvents or ligands.