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Coupling Exonuclease Digestion with Selective Chemical Labeling for Base-resolution Mapping of 5-Hydroxymethylcytosine in Genomic DNA

Aurelien A. Serandour 1, 2 Stéphane Avner 3 Gilles Salbert 3
1 ECN - École Centrale de Nantes
2 CRCINA - Département NOHMAD - Equipe 11 - Integrative oncogenomics of multiple myeloma pathogenesis and progression
CRCINA - Centre de recherche de Cancérologie et d'Immunologie / Nantes - Angers
3 IGDR - Institut de Génétique et Développement de Rennes
Abstract : This protocol is designed to obtain base-resolution information on the level of 5-hydroxymethylcytosine (5hmC) in CpGs without the need for bisulfite modification. It relies on (i) the capture of hydroxymethylated sequences by a procedure known as 'selective chemical labeling' (see Szulwach et al., 2012) and (ii) the digestion of the captured DNA by exonucleases. After Illumina sequencing of the digested DNA fragments, an ad hoc bioinformatic pipeline extracts the information for further downstream analysis.
Keywords : Exonuclease digestion CpG Selective chemical labeling 5-Hydroxymethylcytosine
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https://hal-univ-rennes1.archives-ouvertes.fr/hal-02062038
Contributor : Xavier Chard-Hutchinson <>
Submitted on : Friday, March 8, 2019 - 3:02:43 PM
Last modification on : Thursday, February 27, 2020 - 4:15:48 PM

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UNIV-RENNES1 | IGDR | CNRS | UNAM | UNIV-ANGERS | BIOSIT | UR1-UFR-SVE | CRCINA | UNIV-NANTES | UNIV-RENNES | EC-NANTES

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Aurelien A. Serandour, Stéphane Avner, Gilles Salbert. Coupling Exonuclease Digestion with Selective Chemical Labeling for Base-resolution Mapping of 5-Hydroxymethylcytosine in Genomic DNA. Bio-protocol , Bio-protocol LCC, 2018, 8 (5), ⟨10.21769/BioProtoc.2747⟩. ⟨hal-02062038⟩

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