US 2008/0176758 A1 · Method for the analysis of the methylation status of a nucleic acid

Abstract

The invention relates to a method for the analysis of the methylation status of a nucleic acid. According to the invention, the method comprises following steps: (a) treating the nucleic acid with a chemical reagent or an enzyme containing solution, whereby the base pairing behavior of methylated cytosine bases and/or unmethylated cytosine bases of the nucleic acid are altered such that methylated cytosine bases become distinguishable from unmethylated cytosine bases, (b) hybridizing to the treated nucleic acid an at least one oligonucleotide, (c) ligating the at least one oligonucleotide to itself to form a circular DNA molecule if a particular methylation status is present in the nucleic acid, and (d) amplifying the DNA molecule formed to detect the methylation status of the nucleic acid.

Claims49

Dependent claims

A method for the analysis of the methylation status of a nucleic acid, comprising the following steps:

treating the nucleic acid with a chemical reagent or an enzyme containing solution, whereby the base pairing behavior of methylated cytosine bases and/or unmethylated cytosine bases of the nucleic acid are altered such that methylated cytosine bases become distinguishable from unmethylated cytosine bases, and
hybridizing to the treated nucleic acid an at least one oligonucleotide that is ligatable to itself if a particular methylation status is present in the nucleic acid, and
ligating the at least one oligonucleotide to itself to form a circular DNA molecule if a particular methylation status is present in the nucleic acid, and
amplifying the DNA molecule formed to detect the methylation status of the nucleic acid.

1 more element

An oligonucleotide for analyzing the methylation status of a nucleic acid with a first and second sequence portion which are located adjacent to each other, which comprises

a first sequence portion for hybridizing to the first sequence portion of the treated nucleic acid,
a second sequence portion for hybridizing to the second sequence portion of the treated nucleic acid, and
a third sequence portion which is located between the first and the second sequence portion of the at least one oligonucleotide and which allows for the at least one oligonucleotide to form a circle-like structure when the first and the second sequence portion of the at least one oligonucleotide are hybridized to the first and the second sequence portion of the treated nucleic acid,
characterized in that the first and the second sequence portion of the oligonucleotide only contain adenine, cytosine, and thymine, but no guanine.

1 more element

Dependencies are inferred from the claim text.

Method for the analysis of the methylation status of a nucleic acid

Life of this patent

Method for the analysis of the methylation status of a nucleic acid

Life of this patent

Abstract

Claims49