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Protocols
Biochemistry
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Coupling Exonuclease Digestion with Selective Chemical Labeling for Base-resolution Mapping of 5-Hydroxymethylcytosine in Genomic DNA
Aurélien A. Sérandour
SA
Stéphane Avner
GS
Gilles Salbert*
0 Q&A
6519 Views
Mar 5, 2018
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.
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