This laboratory project is one component of a semester-long advanced biochemistry laboratory course that uses several complementary techniques to study tRNA[superscript Phe] conformational changes induced by ligand binding. In this article we describe a set of experiments in which students assay metal-catalyzed hydrolysis of tRNA[superscript Phe] using gel electrophoresis. tRNA[superscript Phe] cleavage patterns are monitored in the presence of lead(II) and other metals cations. Small molecule ligands are also studied to determine their ability to enhance or inhibit lead(II)-catalyzed cleavage of tRNA[superscript Phe]. Gel bands are quantified to determine ligand-binding equilibrium constants for the small molecule ligand-tRNA[superscript Phe] adducts. By pooling class data from the metal and small molecule ligand experiments, students draw conclusions regarding how metal-catalyzed hydrolysis is affected by various parameters such as tRNA[superscript Phe] structure, ligand charge, and inorganic chemistry of the metal ion complexes. (Contains 2 tables, 1 figure and 2 notes.)