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. 2017 Mar;23(3):355–364. doi: 10.1261/rna.059824.116

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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 6. — Similar to glmS^WT and glmS^AAA, glmS^Ca RNA cleavage proceeds by internal transesterification, but at a site 1 nt distal. (A) 5′ Cleavage product end-group analysis. 5′ ³²P-radiolabeled substrate was cleaved in trans with the three ribozymes for 30 min and treated with acid or with phage T4 polynucleotide kinase prior to analysis by denaturing PAGE, as previously described (Winkler et al. 2004; Xiao et al. 2008; Lau and Ferré-D'Amaré 2013). (B) Mapping of glmS^Ca cleavage site. 5′ ³²P-radiolabeled of RNA substrates of varying lengths with or without single-site 2′-deoxy substitutions were cleaved by the three ribozymes and analyzed by denaturing PAGE. (Hy.) Hydrolysis ladder.