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Nat Methods. 2011 Nov 20;9(1):75-7. doi: 10.1038/nmeth.1779.

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

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  • 1Department of Chemistry and Institute for Biophysical Dynamics, The University of Chicago, Chicago, Illinois, USA.

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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PMID:
22101853
[PubMed - indexed for MEDLINE]
PMCID:
PMC3646335
Free PMC Article
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