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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Methods Enzymol. 2010;472:153–178. doi: 10.1016/S0076-6879(10)72011-5

Figure 1. Capturing the conformational dynamics of single pre-mRNA molecules through smFRET in real-time.

Figure 1

(A) A pre-mRNA molecule is immobilized through a 2'-O-methylated capture oligonucleotide, and is bound to a PEG passivated quartz slide through a biotin-streptavidin interaction. The inset depicts a portion of a field of view captured by the I-CCD camera, and how the donor (Cy3) and acceptor (Cy5) fluorophores can be captured simultaneously. Peak finder algorithms are used to automatically find and match corresponding fluorophores from single molecules (white circles). (B) Exemplary fluorescence intensity and FRET changes of a single molecule.