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. 2007 Jan 31;35(4):1270–1278. doi: 10.1093/nar/gkl1151

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© 2007 The Author(s).

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — (A) Autoradiography of the 3’-end labeled D. hafniense tRNA^Pyl transcripts after enzymatic cleavage with RNase S1 (S1) and RNase T1 (T1) in the presence (+) and absence (−) of D. hafniense PylRS. D is Decade Marker (Ambion) that contains ³²P-labeled RNA oligos whose nucleotide length is indicated on the left. C is untreated tRNA. RNase S1 (S1) cuts in single stranded regions, and no protection was observed in the anticodon stem. RNase T1 (T1) cuts at single stranded G and protection was observed at G14 and G48. (B) Autoradiography of the 3'-end labeled, phosphorothioate-containing D. hafniense tRNA^Pyl transcripts after cleavage by iodine in the presence and absence of D. hafniense PylRS. The footprinting experiments were performed using transcripts statistically substituted with ATP[αS] (A lanes), UTP[αS] (U lanes), CTP[αS] (C lanes), and GTP[αS] (G lanes). Lanes are numbered to indicate the PylRS concentration: (1) no PylRS, (2) 28 μM PylRS, (3) 14 μM PylRS, and (4) 7 μM PylRS.