Workflow from Scientific Research

Schematic workflow of the G4switch-biotin Chem-map in U2OS cells. In permeabilized cells with or without 405 nm illumination, a precomplex comprising biotinylated 9 (red orange and blue) and anti-biotin antibody (grey) binds to DNA targets of 9, followed by a sequential secondary antibody (purple) binding and a fusion transposome pA-Tn5 (light blue) binding to the target sites. Mg 2+ was then used to activate the Tn5 transposase in situ for DNA cleavage and sequencing adapter (orange and teal) integration adjacent to the 9 binding sites. After DNA isolation, PCR amplification of the fragments labelled with adapters at both ends generated the library, which was sequenced to identify the genomic binding sites of 9.

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