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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 5. — 5′-³²P-labeling of random alkali fragments (A), and 2D-TLC analysis of the wobble base nucleotide in U34 mutants of H. marismortui tRNA₂^Ile (B) and tRNA₁^Ile (C). Purified tRNAs were cleaved randomly with alkali; fragments were 5′-end labeled with ³²P and separated by gel electrophoresis. The 5′-end nucleotide of the fragment corresponding to U*34 was analyzed by nuclease P1 digestion and 2D-TLC. Nonradioactive markers pA, pC, pG, and pU were visualized by UV-shadowing and are indicated by circles. (pU*) position of cnm⁵U.