Skip to main content
. 2016 Mar 28;113(16):4338–4343. doi: 10.1073/pnas.1600223113

Fig. 2.

Fig. 2.

Dual-labeling of 5mC and 5hmC for simultaneous imaging. (A) General procedure for dual-labeling and simultaneous imaging of 5mC and 5hmC. DNA fragments are end-labeled with biotin, 5hmC is labeled with Cy5, and 5mC is labeled with Cy3. The labeled DNA is immobilized to the microscope slide and imaged with single-molecule TIRF microscopy. (B) Fluorophore counts of dual-labeled P60 and P14 mouse cerebellum genomic DNA with example images shown on the right. (C) Distribution of multiple fluorophores on DNA fragment of mouse cerebellum when detecting 5mC. (D) The 5mC to 5hmC ratio in mouse cerebellum genomic DNA after adjusting multiple fluorophores. (E) The 5mC level of mouse cerebellum DNA. Error bars, mean ± SD (n = 15 counting regions).