Patent ID: 8105782
Filing Date: 2012-01-31
Classification: C12Q

Abstract:
1. A method of determining a methylation profile for a target nucleic acid sequence, said method comprising (a) treating the nucleic acid sequence with a bisulfite, to convert unmethylated cytosines within the nucleic acid sequence to uracil thereby creating a converted nucleic acid sequence; (b) amplifying said converted nucleic acid sequence using oligonucleotide primers specific only for said converted nucleic acid sequence, wherein said primers do not discriminate between methylated and unmethylated alleles, said amplification conditions including the incorporation of a signal agent in the amplified nucleic acid, said signal agent being a double-stranded DNA dye capable of providing a distinction between double-stranded and single-stranded nucleic acid; and then, following steps (a) and (b), (c) detecting the amount of said signal agent during transition of the amplified nucleic acid between double-stranded and single-stranded as a result of a temperature change; (d) determining the methylation profile of said target nucleic acid sequence by measuring the amount of signal agent detected during said temperature change, wherein said determining step comprises resolving a heterogeneous methylation pattern, if present, in said target nucleic acid sequence.