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. 2009 Mar 20;37(9):3061–3073. doi: 10.1093/nar/gkp182

Figure 6.

Figure 6.

Run-off sequencing of purified cleavage products of I-SceI, CdnDI and the hybrid endonuclease with the 10S linker. Each cleavage product was sequenced toward the cleavage site. The sharp drop-off after a particular base indicates the end of the fragment, and thus the site of DNA cleavage. (a) I-SceI. The fragment ends generated by I-SceI are in agreement with its previously reported internal cleavage site (62). (b) CdnDI. The overlapping fragments generated by CdnDI indicate 5′, four-base overhangs resulting from cuts at positions 2/6 relative to the I-SceI recognition site. The asterisks denote non-templated additions of ‘A’, as discussed in ‘Results’ section. CdnDII and the hybrid endonuclease with the 20S linker produced run-off sequencing results indistinguishable from those for CdnDI. (c) The hybrid enzyme with the 10S linker. The overlapping fragments generated by this enzyme indicate 5′, four-base overhangs resulting from cuts at positions 1/5 relative to the I-SceI recognition site.