Supplementary Figure 4: Properties of 5fC labeling products during PCR and/or bisulfite treatment.
From: Bisulfite-free, base-resolution analysis of 5-formylcytosine at the genome scale
(a) 5fC labeling with diethyl malonate protects the product from bisulfite-mediated deamination and hence is read as C in canonical bisulfite sequencing, similar to the previously reported fCAB-Seq8. (b) The adduct between 5fC and 1,3-indandione results in C-to-T transition during PCR. (c) Single-nucleotide incorporation of dATP, dGTP, dCTP and dTTP opposite the 5fC-I for 1, 2, 5, 10, 30 and 60 min, respectively. A template where X is a T was also included as a positive control. (d) Quantification of single-nucleotide incorporation. Values are presented as mean ± s.d. (n = 3). (e) Schematic comparisons of fC-CET with fCAB-Seq.
