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. Author manuscript; available in PMC: 2016 May 7.
Published in final edited form as: Cell. 2015 Apr 30;161(4):879–892. doi: 10.1016/j.cell.2015.04.010

Figure 2. Schematic Diagram of 6mA-IP-seq and 6mA-CLIP-exo.

Figure 2

For 6mA-IP-seq (left), fragmented genomic DNA (gDNA) is ligated to a Y-shaped adapter with specific index sequence, denatured, and immunoprecipitated using anti-6mA antibody. The captured DNA is eluted with 6mA single nucleotide, and PCR amplified to construct the DNA library. Simultaneously, the input library was obtained from the ligated DNA before immunoprecipitation. For 6mA-CLIP-exo (right), fragmented gDNA is incubated with anti-6mA antibody, crosslinked by 254 nm UV irritation, and immunoprecipitated. The crosslinked DNA is ligated to adapter R1 on beads, followed by 5′ to 3′ exonuclease digestion. Antibody-protected DNA is preserved, and a 2nd-strand DNA synthesis is performed after protease digestion of the antibody. A second ligation to adapter R2 provides the template for PCR amplification to construct the library for high-throughput sequencing. Boundaries were determined by the sequencing ends of the 6mA-CLIP-exo-seq to provide a high resolution localization of 6mA.

See also Figure S2.