Single-Molecule Detection of 5-Hydroxymethylcytosine in DNA through Chemical Modification and Nanopore Analysis


  • We thank Oxford Nanopore Technology for financial support.


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DNA threading the needle: A new method of single-molecule detection of 5-hydroxymethylcytosine (5hmC) in DNA has been developed. Selective thiol substitution of 5hmC (giving SMC) in a single-step, bisulfite-mediated reaction (see scheme) allows the incorporation of a peptide (yellow sphere) or biotin into DNA. Modified 5hmC bases can be readily distinguished at the single-molecule level using protein nanopore analysis.