nmr spectroscopy is a technique that has been used over the years to study important dynamic aspects of base flipping. this technique allows researchers to determine the physical and chemical properties of atoms and other molecules by utilizing the magnetic properties of atomic nuclei. in addition, nmr can provide a variety of information including structure, reaction states, chemical environment of the molecules, and dynamics. during the dna base flipping discovery experiment, researchers utilized nmr spectroscopy to investigate the enzyme-induced base flipping of hhai methyltransferase. in order to accomplish this experiment, two 5-fluorocytosine residues were incorporated into the target and the reference position with the dna substrate so the f chemical shift analysis could be performed. once the f chemical shift analysis was evaluated, it was then concluded that the dna complexes existed with multiple forms of the target 5-fluorocytosine along the base flipping pathway.