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. 2015 May;21(5):1031–1041. doi: 10.1261/rna.047845.114

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© 2015 Rodgers et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 2. — SNAP-SiMPull characterization of interactions between RNAs and immobilized BBP (A) Cartoon representation of the single-molecule assay for immobilized DY649 BBP-SNAP_AP binding to a Cy3-labeled oligo containing the UACUAAC sequence. (B) Representative Cy3 fluorescence trace illustrating many binding events of Cy3-labeled RNAs to a single molecule of BBP-SNAP_AP. (C) Cartoon representation of the single-molecule assay for immobilized DY649 BBP-SNAP_AP binding to a Cy3-labeled oligo that does not contain a branchsite sequence (control oligo). (D) Representative Cy3 fluorescence trace illustrating few binding events of Cy3-labeled RNAs lacking a branchsite sequence to BBP-SNAP_AP. (E) Quantification of the appearance of Cy3-RNA fluorescent spots in the presence and absence of immobilized BBP-SNAP_AP. Error bars were calculated using the standard error of the mean. In the absence of BBP-SNAP_AP pull-down, approximately sevenfold fewer Cy3-RNA spots appeared in randomly selected areas of interest (AOIs).