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Updated: Dec 11, 2025

Selective Capture of 5-hydroxymethylcytosine from Genomic DNA
Published on: October 5, 2012
Selective Chemical Labeling and Sequencing of 5-Carboxylcytosine in DNA at Single-Base Resolution
Yalun Xie1, Yafen Wang1, Zhiyong He1
1College of Chemistry and Molecular Sciences, Key Laboratory of Biomedical Polymers of Ministry of Education, the Institute for Advanced Studies, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University, Wuhan, Hubei 430072, P. R. China.
Abstract:
5-Carboxylcytosine (5caC) plays a vital role in the dynamics of DNA demethylation, and sequencing of its sites will help us dig out more biological functions of 5caC. Herein, we present a novel chemical method to efficiently label 5caC distinguished from other bases in DNA. Combined with bisulfite sequencing, 5caC sites can be located at single-base resolution, and the efficiency of 5caC labeling is 92% based on the Sanger sequencing data. Furthermore, dot blot assays have confirmed that 5caC-containing DNA isolated from HeLa cells was successfully labeled using our method. We expect that our strategy can be further applied to selectively tagging other carboxyl-modified bases and mapping their sites in RNA.
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