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

Song, Chun-Xiao; Clark, Tyson A; Lu, Xing-Yu; Kislyuk, Andrey; Dai, Qing; Turner, Stephen W; He, Chuan; Korlach, Jonas · Nat Methods · 2011

basic_science · Level V

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Abstract

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.

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