Abstract
Nucleic acid melt data is ubiquitous, whether obtained by ultraviolet-visible spectroscopy (UV-vis), fluorescence from Förster Resonance Energy Transfer (FRET) pairs, intercalating dyes, or other reporters, and, in the context of quantitative Polymerase Chain Reaction (qPCR) experiments and High-Resolution Melting (HRM) experiments, plentiful. While van’t Hoff analysis of nucleic acid melting data is considered to be straightforward, considerable care is needed to ensure that it yields precise and accurate thermodynamic parameters which are consistent with other measurements and useful for prediction.
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