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

Authors :: Song CX
Clark TA
Lu XY
Kislyuk A
Dai Q
Turner SW
He C
Korlach J
Source :: Nature methods [Nat Methods] 2011 Nov 20; Vol. 9 (1), pp. 75-7. Date of Electronic Publication: 2011 Nov 20.
Publication Year :: 2011
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.