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  Home > JCE Print > Journal of Chemical Education > Issues > 2008  > May  >
In the Laboratory
UV Thermal Melting Curves of tRNAPhe in the Presence of Ligands
Sarah R. Kirk, Todd P. Silverstein, and Karen L. McFarlane Holman
Department of Chemistry, Willamette University, Salem, OR 97301
Cover
May 2008
Vol. 85 No. 5
p. 674

Abstract
This laboratory project is one component of a semester-long advanced biochemistry laboratory course that uses several complementary techniques to study tRNAPhe conformational chvanges induced by ligand binding. In this article we describe a set of experiments in which the thermal unfolding of tRNAPhe is studied with UV–vis spectrophotometry. Over the course of two laboratory periods, students examine the effects of magnesium and small molecule ligands on tRNAPhe conformational stability. The data collected in this laboratory are fit to saturation binding curves, the ligand-binding equilibrium constants are determined, and conclusions are drawn about tRNAPhe ligand binding modes and the impact of various types of ligands on tRNA structure.
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Detailed student manual; Instructor notes
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Citation
Kirk, Sarah R.; Silverstein, Todd P.; Holman, Karen L. McFarlane. J. Chem. Educ. 2008, 85, 674.
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Keywords
Biochemistry; Bioinorganic Chemistry; Bioorganic Chemistry; Biophysical Chemistry; Hands-On Learning / Manipulatives; Laboratory Instruction; Nucleic Acids / DNA / RNA; Thermal Analysis; Upper-Division Undergraduate; UV-Vis Spectroscopy
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History
Created:
Last Updated:
3/25/2008
3/28/2008
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Experiments, laboratory exercises, lecture demonstrations, and other descriptions of the use of chemicals, apparatus, instruments, computers, and computer interfaces are presented in the Journal of Chemical Education as illustrative of new or improved ideas or concepts in chemistry instruction and are directed at qualified teachers. Although every effort is made to assure and encourage safe practices and safe use of chemicals, the Journal of Chemical Education cannot assume responsibility for uses made of its published materials. Many chemicals are hazardous. Precautions for the safe use of hazardous chemicals and directions for their proper disposal are described in the Material Safety Data Sheets and on the labels. We strongly urge all those planning to use materials from our pages to make choices and to develop procedures for laboratory and classroom safety in accordance with local needs and situations.
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