Method for detecting methylation status by using methylation-independent primers

A method for detecting the methylation status of a CpG-containing nucleic acid in a sample comprising the steps of:

• a) providing said sample comprising a CpG-containing nucleic acid,
• b) denaturing said CpG-containing nucleic acid, thereby producing denatured CpG-containing nucleic acid,
• c) treating said denatured CpG-containing nucleic acid with bisulphite which modifies at least one unmethylated cytosine in said CpG-containing nucleic acid, thereby producing treated CpG-containing nucleic acid,
• d) amplifying said treated CpG-containing nucleic acid by means of at least one methylation-independent oligonucleotide primer, which comprises within the first 10 nucleotides of the 5′-end, at least one CpG dinucleotide in a region which hybridizes to the CpG-containing nucleic acid, wherein said at least one methylation-independent oligonucleotide primer hybridizes to both methylated and unmethylated CpG-containing nucleic acids after modification, and
• e) analyzing the amplified CpG-containing nucleic acid by melting curve analysis.