Figure 3

(a) Steps of the labeling scheme (see Fig. 2a for description). (b–d) Denaturing gel analyses of 34 nt DNA constructs featuring either 5mC (b) or 5hmC (c–d) at base position 22 and labeled using WT TDG following oxidation of each base with TET. In (d), a treatment with βGT prevents labeling of 5hmC specifically. Lane (i): annealed oligonucleotide ± βGT; lane (ii): following glycosylase/endonuclease treatment; lane (iii): following polymerase fill-in with a biotinylated nucleotide to yield a labeled product (red). Construct lengths at left apply to all gels and * indicates DNA length plus biotin tag. Directly below lanes (ii) and (iii) are listed target product yields from the previous step followed by the net yield in red. Full gels are shown in Supplementary Figure S3.