The discovery of 5-methylcytosine oxidative products led to the development of a large number of techniques to allow the detection of cytosine modifications and to map them genome-wide. Here is described an antibody-based method to detect 5-hydroxymethylcytosine (5-hmC) in mammalian cells. This protocol consists of an extraction of genomic DNA followed by a three-day immunoprecipitation procedure. 5-hmC containing immunoprecipitated DNA fragments can be further analysed by qPCR or submitted to high throughput sequencing. It is important to note that this approach allows the mapping of 5-hmC only at low resolution and, as a consequence, other techniques must be used for single nucleotide resolution.
Institut de Génétique et Développement de Rennes
Université de Rennes 1
263 Avenue Général Leclerc
Corresponding author: Gilles Salbert