Figure 7:

Detection of Tet1-mediated 5mC oxidation. (A) Detection of Tet oxidation activity using 20 bp oligonucleotides. In all, 0.5 µM of fully methylated 20 bp DNA was incubated with 2 µM of Tet1CD. After 2 h incubation, DNA was used for HRM analysis. Independent experiments were repeated twice. Shown is one representative result with three technical replicates. The P-values of student’s t-test are indicated in the plot. (B) Detection of Tet oxidation activity using a PCR fragment containing multiple methylated cytosines. In all, 50 ng of 377 bp methylated PCR fragment was incubated with 2 µM of Tet1CD. After 2 h incubation, DNA was used for HRM analysis. Independent experiments were repeated twice. Shown is one representative result with four technical replicates. hmC: 5-hydroxymethylcytosine; fC: 5-Formylcytosine; caC: 5-carboxylcytosine.