Single-Molecule Multicolor FRET Assay for Studying Structural Dynamics of Biomolecules.

Over the last 2 decades, single-molecule Forster resonance energy transfer (FRET) has been widely used to address important questions in molecular biology. However, a conventional approach based on a single donor-acceptor pair is not powerful enough to study complex biological systems. To address this challenge, single-molecule multicolor FRET techniques have been developed. In this chapter, we present practical considerations required for the successful implementation of single-molecule multicolor FRET in the laboratory.