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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Nat Methods. 2013 Mar 17;10(4):361–365. doi: 10.1038/nmeth.2408

Figure 1.

Figure 1

BLESS workflow and specificity. (a) DSBs are ligated in situ to a proximal linker (red arch) covalently linked to biotin (orange oval) (1), gDNA is extracted and fragmented (2), and labeled fragments are captured on streptavidin beads (gray ovals) (3). A distal linker (blue arch) is then ligated to the free extremity of captured fragments (4), and fragments are released by linker digestion with I-SceI (5). Released fragments are amplified by PCR using linker-specific primers (6), and sequenced (7). (b) Structure of linkers. Both proximal (P) and distal (D) linkers share an XhoI site (yellow), the I-SceI endonuclease minimal recognition site (non-highlighted letters), and a seven-thymine loop (bold). Each linker contains a specific barcode sequence marking the ligation site (orange and brown). The proximal linker is biotinylated (orange oval). (c) Proportion of fragments with proximal (P) and distal (D) barcodes in single-end (SE) and pair-end (PE) Illumina sequencing experiments. Mean ± s.d. is shown.