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. 2012 Jan 28;40(10):4589–4603. doi: 10.1093/nar/gks006

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© The Author(s) 2012. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Representative Northern blots showing the determination of the percentage of aminoacylated tRNA^Arg2 and tRNA^Pro. Total RNA isolated from SK10153 (wild-type), MG1693 (rph-1), SK10592 (Δrnt), SK10148 (Δrnt rph-1) and SK10020 (Δrnt ΔpcnB rph-1) were untreated (lanes 1–5) or treated with 0.5 M Tris (pH 9) to chemically deacylate tRNAs (lanes 6–10) and were separated using acid urea polyacrylamide gel as described in Materials and Methods. The position marked with I indicated uncharged form of tRNA. The position marked with II indicated either charged and/or high molecular weight immature tRNAs [with or without poly(A) tails]. The net extent of aminoacylation (lanes 1–5) for band II was calculated as percentage of total counts (bands I + II) minus the percentage of high molecular weight species of total counts (bands I + II) after Tris treatment in lanes 6–10 (33). The aminoacylation levels of all tRNAs studied are reported in Table 4.