Labeling DNA (or RNA) for single-molecule FRET

Chirlmin Joo; Taekjip Ha

doi:10.1101/pdb.prot071027

Labeling DNA (or RNA) for single-molecule FRET

Chirlmin Joo, Taekjip Ha

Department of Physics

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

Single-molecule (sm) fluorescence detection is a powerful method for studying biological events without time and population averaging. Förster (fluorescence) resonance energy transfer (FRET) is a spectroscopic technique for measuring distances in the 30-80 Å range in which excitation energy of a donor molecule is transferred to an acceptor via interaction between two induced dipoles. A variant of smFRET is based on total internal reflection (TIR) microscopy. This protocol describes the labeling of nucleic acids (DNA or RNA) for smFRET with TIR microscopy. The protocol is optimized for 5 nanomoles of DNA that has an amine-modified thymine in its sequence. It is a simplified version of a protocol from Invitrogen.

Original language	English
Pages (from-to)	1005-1008
Number of pages	4
Journal	Cold Spring Harbor Protocols
Volume	7
Issue number	9
DOIs	https://doi.org/10.1101/pdb.prot071027
State	Published - Sep 2012

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Labeling DNA (or RNA) for single-molecule FRET

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