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. 2009 Dec 2;38(4):1294–1303. doi: 10.1093/nar/gkp1092

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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 5. — Run-off sequencing of nicked products by BtsI nicking variants. The cleavage product of mutants E128F (A) and D388A/E403A/E405A (B) were gel-purified and subjected to Sanger sequencing on both strands. The recognition sequence is bolded and the cleavage site was indicated by arrows. The extra A peak at the end of run-off sequence was added by the template independent terminal transferase activity of Taq DNA polymerase during sequencing. The extra A peak and drop off in peak signal indicate the nicking site (cleaved template).