Patent ID: 7364855

Claim:
A method for determining the degree of methylation of at least one target nucleotide in at least one target nucleic acid sequence, comprising: forming a misligation reaction composition comprising (a) the at least one target nucleic acid sequence, (b) at least one ligation probe set comprising at least one first probe and at least one second probe, wherein the at least one first probe comprises at least one first target-specific portion with at least one nucleotide mismatches the target nucleotide sequence and the at least one second probe comprises at least one second target-specific portion wherein at least one nucleotide mismatches the target nucleotide sequence, wherein the two portions are adjacent to one another and (c) at least one ligation agent; subjecting the misligation reaction composition to at least one cycle of ligation to generate at least one misligation product; and determining the degree of methylation of the at least one target nucleotide.