Detection of 5-Hydroxymethylcytosine in DNA by Transferring a Keto-Glucose by Using T4 Phage β-Glucosyltransferase

Capture with ketone: 5‐Hydroxylmethylcytosine (5‐hmC) in DNA can be selectively labeled with a keto‐glucose and subsequently linked to biotin via a ketoxime linker for enrichment and detection/sequencing purposes (see scheme). Keto‐glucose can be more efficiently transferred than azide‐glucose by β‐glucosyltransferase, and can provide single‐base resolution detection/ sequencing of 5‐hmC.