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. 2014 Feb;20(2):177–188. doi: 10.1261/rna.042358.113

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© 2014 Mandal 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 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

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FIGURE 3. — State of in vivo aminoacylation of H. marismortui wild-type and U34 mutant tRNA₂^Ile (A) and wild-type and U34 mutant H. marismortui tRNA₁^Ile (B). Total tRNA was isolated under acidic conditions and analyzed by acid urea PAGE followed by Northern hybridization using probes specific for H. marismortui tRNA₂^Ile (A, top panel) and H. marismortui tRNA₁^Ile (B, top panel), respectively. The same blots were stripped and rehybridized using probes specific for the endogenous H. volcanii tRNA₂^Ile (A, bottom panel) and H. volcanii tRNA₁^Ile (B, bottom panel). Note that the H. volcanii tRNA₁^Ile-specific probe also picks up the overexpressed H. marismortui tRNA₁^Ile because of the high levels of overexpression of H. marismortui tRNAs and close sequence similarity between the overexpressed and the endogenous tRNA₁^Ile. (ac) tRNA isolated under acidic conditions; (OH⁻) tRNA after deacylation by base-treatment.