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Abstract
Described herein is a multiplex 5mC marker barcode counting (MMBC) to quantify 5mC markers in DNA, in which multiple 5mC markers could be targeted for capture and linear amplification. The inventors' method provides highly sensitive and specific quantification of multiple 5mC loci. Accordingly, aspects of the disclosure relate to a method for detecting methylated or unmethylated cytosines in one or more regions of target nucleic acids, the method comprising i) combining a solution comprising the target nucleic acids with a deaminating agent to convert unmethylated cytosines in the target nucleic acids to uracils; ii) next contacting the solution with at least two probes under conditions that allow for the hybridization of the two probes to one target nucleic acid region; wherein a terminal end from each probe hybridizes adjacently to the target nucleic acid region; iii) contacting the solution comprising the hybridized probes and target nucleic acids with a ligase under conditions that allow for the ligation of the terminal ends of the adjacently hybridized probes; and iv) detecting the adjacently hybridized ligated probes in the solution.