Abstract
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.
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