This seminar in the JASCO series on Molecular Spectroscopy takes a closer look at bio-analysis using temperature control and thermal melting. This technique is a powerful tool to monitor protein stability and protein-protein/protein-nucleic acid binding interactions and is used with a range of spectroscopic techniques including fluorescence, UV-Visible, and circular dichroism.
Although the webinar focuses on fluorescence and the complimentary technique circular dichroism spectroscopy for the structural analysis of proteins, principles can also be extended to structural changes in nucleic acids (DNA and RNA). This webinar will be valuable for researchers who are new to thermal melt studies, as well as those more experienced, because it provides practical details on how to get the best data and results, including design, set-up, collection, and analysis.
Key points include:
- Experimental design, including optimization of measurement parameters to achieve high quality data, such as
- Single-point vs. spectral data
- Staged temperature profiles for efficiency and accuracy
- Ramping vs. equilibrating temperature
- Analysis and data modeling, the pitfalls of poor data
- Relevant examples will be provided to illustrate good practices and highlight what kind of information can be gained
Understanding, Analyzing & Optimizing Thermal Melts for Fluorescence, UV & CD Applications