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. 2014 Sep 8;42(17):11144–11155. doi: 10.1093/nar/gku797

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Cleavage of ssDNA by Pcal_0546 and SSO1391. (A, B) The 3′- or 5′-[³²P]-labeled ssDNA was incubated without protein (lane C) or with indicated amounts of Pcal_0546 (A) or SSO1391 (B) for 15 minutes at 45ºC and analyzed by denaturing gel electrophoresis. (C, D) Endonuclease assays: the M13mp18 ssDNA (5 nM) was incubated at 45ºC without protein (lane C) or in the presence of indicated amounts of Pcal_0546 (C) or SSO1391 (D) and the reaction products were analyzed by agarose gel electrophoresis and SYBR Green staining. (E) Mass spectrometry analysis of the ssDNA cleavage products generated by Pcal_0546 and SSO1391. Relative abundance of ions and retention times corresponding to dC (m/z 226.0834) and dCMP (m/z 306.0497) are shown for standards (black) and products of Pcal_0546 or SSO1391 mediated cleavage of a 10 nt long oligonucleotide (gray).