Department of Chemistry and Institute for Biophysical Dynamics, The University of Chicago, Chicago, Illinois, USA.
Nat Methods. 2011 Nov 20;9(1):75-7. doi: 10.1038/nmeth.1779.
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.
我们描述了一种具有单分子灵敏度的、用于基因组 DNA 中 5-羟甲基胞嘧啶（5-hmC）的链特异性、碱基分辨率检测方法，该方法结合了 5-hmC 的生物正交、选择性化学标记方法和单分子实时（SMRT）DNA 测序。该化学标记不仅允许富含 5-hmC 的 DNA 片段进行亲和富集，而且还增强了 SMRT 测序过程中 5-hmC 的动力学信号。我们应用该方法在高置信度的基因组 DNA 样本中对 5-hmC 进行了测序。
