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. 2010 Nov 8;39(2):e11. doi: 10.1093/nar/gkq1082

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Published by Oxford University Press 2010.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Chemiluminescence-based Assay. (A) Schematic representation of the flap DNA substrate; flap length is indicated by the number in italics. The three deoxyoligonucleotide sequences are shown. The biotin and fluorescein (FITC) labels introduced for bead recognition are indicated on the 5′-ends of the flap and upstream strands, respectively. The FEN1 cleavage site is indicated by the broken arrow. Upon red-shifted light excitation (λ₆₈₀) of the donor bead, singlet oxygen is generated. If singlet oxygen encounters an acceptor bead within its traveling range, it triggers the emission of blue-shifted light from the acceptor bead (λ_520–620). (B) Substrate titration against beads (N = 2). A strong dose-dependent signal associated with a ‘hook-shaped’ curve was observed as expected (see text for details), in contrast to the unlabeled control (empty triangles), which elicited only a background signal. (C) Concentration dependence of the cleavage reaction on FEN1 protein (N = 2).