5fC Chemical Labeling and C-to-T Transition During PCR

fC-CET is a bisulfite-free method for whole-genome analysis of 5fC. The method is based on selective chemical labeling of 5fC and subsequent C-to-T transition during PCR (Xia et al., 2015). In this method, gDNA is labeled sequentially with an azido derivative of 1,3-indandione (AI) and then conjugated to biotin using click chemistry. The gDNA is enriched by streptavidin pull-down and ligated to adapters, followed by PCR and sequencing. C-to-T transitions during PCR amplification are searched specifically to define 5fC sites in the genome.


  • Results are in agreement with fC-Seal
  • Sequences with a single 5fC are enriched ~100-fold
  • No noticeable DNA degradation; ideal for analyses of precious DNA samples



Illumina Library prep and Array Kit Selector


None available yet


Xia B., Han D., Lu X., et al. Bisulfite-free, base-resolution analysis of 5-formylcytosine at the genome scale. Nat Methods. 2015;12:1047-1050