TAmC-Seq
TAmC-Seq
TAmC-Seq selectively tags 5-methylcytosine with an azide residue that can be further labelled with a biotin for affinity purification.
In this method the endogenous 5hmC in genomic DNA is protected by glycosylation with regular glucose. The 5mC residues are converted to 5hmC via mTet1-catalysed oxidation. The 5hmC is then labelled via b-GT-mediated glucosylation with a modified glucose moiety (6-N3-Glucose) to generate 6-N3-b-glucosyl-5-hydroxymethyl-cytosine (N3-5gmC), which can be labelled using click chemistry for detection, affinity purification and sequencing.