Poster abstracts
Poster number 112 submitted by William Roy
Understanding the complications of working with G-quadruplex systems in the single molecule FRET assay
William A. Roy, Jr. (Department of Physics, Kent State University), Parastoo Maleki (Department of Physics, Kent State University), Jagat B. Budhathoki (Department of Physics, Kent State University), Hamza Balci (Department of Physics, Kent State University)
Abstract:
We present our studies on the methodology aspect of applying single molecule Förster Resonance Energy Transfer (smFRET) to study G-quadruplex (GQ) systems. These non-canonical nucleic acid structures have proved to be challenging systems to work with at the single molecule level due to their unusual sensitivity to environmental variables and the preparation protocol. Unless thoroughly understood, these sensitivities may lead to misinterpretation of the data and in inconsistent outcomes in measurements. The annealing and storage conditions, the location of fluorophores on the DNA construct with respect to GQ, and the ionic conditions of the experiment are some of the factors that are of critical importance for the outcome of measurements. Some of these factors are well understood and methods to resolve them have been developed in bulk assays, while others are unique for single molecule assays. We will summarize the knowledge and best practices learned from bulk or single molecule measurements that will help address these issues and result in more reliable smFRET data on GQ-involving systems.
Keywords: g-quadruplex, single molecule, FRET