Sensitive and specific single-molecule sequencing of 5-hydroxymethylcytosine.

Authors: Song, Chun-Xiao and Clark, Tyson A and Lu, Xing-Yu and Kislyuk, Andrey and Dai, Qing and Turner, Stephen W and He, Chuan and Korlach, Jonas

We describe strand-specific, base-resolution detection of 5-hydroxymethylcytosine (5-hmC) in genomic DNA with single-molecule sensitivity, combining a bioorthogonal, selective chemical labeling method of 5-hmC with single-molecule, real-time (SMRT) DNA sequencing. The chemical labeling not only allows affinity enrichment of 5-hmC-containing DNA fragments but also enhances the kinetic signal of 5-hmC during SMRT sequencing. We applied the approach to sequence 5-hmC in a genomic DNA sample with high confidence.

Journal: Nature methods
DOI: 10.1038/nmeth.1779
Year: 2012

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