METHYLATED CPG ISLAND AMPLIFICATION (MCA)

摘要

A method is provided for identifying a methylated CpG containing nucleic aci d including contacting a nucleic acid with a methylation sensitive restriction endonuclease that cleaves unmethylated CpG sites, contacting the sample with an isoschizomer of the methylation sensitive restriction endonuclease, where in the isoschizomer cleaves both methylated and unmethylated CpG sites. The method also includes adding an oligonucleotide under conditions and for a ti me to allow ligation of the oligonucleotide to nucleic acid cleaved by the restriction endonuclease, and amplifying the nucleic acid. A method is also provided for detecting an age associated disorder including contacting a nucleic acid with a methylation sensitive restriction endonuclease that cleaves unmethylated CpG sites, contacting the sample with an isoschizomer o f the methylation sensitive restriction endonuclease, wherein the isoschizomer cleaves both methylated and unmethylated CpG sites. The method also includes adding an oligonucleotide under conditions and for a time to allow ligation of the oligonucleotide to nucleic acid cleaved by the restriction endonuclease, and amplifying the nucleic acid. The amplified digested nucleic acid is adhered to a membrane, and hybridized with a probe of interest. A kit useful for detection ofa CpG containing nucleic acid is also provided.