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. 2009 Dec 29;38(6):1950–1963. doi: 10.1093/nar/gkp1190

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© The Author(s) 2009. 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/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Stalled ΔH RNAPs can efficiently elongate to the run-off. Ternary complexes were stalled at +20, washed and the reactions divided into two aliquots. One aliquot was untreated (lane a of each panel), the other was chased by the addition of a complete set of non-labeled NTPs for 3 min at 70°C (lane b in each panel), and both were then analyzed on a 20% polyacrylamide gel. In lanes 1, 2 and 4, the ability of the ΔH and wt enzyme to synthesize a 20-nt stalled transcript in the presence and absence of subunit H was analyzed. Reactions were chased by the addition of a complete set of NTPs not containing radioactivity and the synthesis of the 145-nt run-off product was analyzed. The RNA products of intermediate size are caused by specific pausing sites for RNAP. The quantification below shows the mean value of the transcriptional activity of the transcript in ternary complexes stalled at position +20 relative to the 145-nt run-off product.