Abstract: A method is described for the detection of the degree of methylation of a specific cytosine in the sequence context 5?-CpG-3? of a genomic DNA sample. In the first step, the genomic DNA is chemically treated in such a way that the cytosine bases are converted to uracil, but not the 5-methylcytosine bases. Then segments of the genomic DNA which contain the said specific cytosine are amplified, whereby the amplified products are given a detectable label and in the following steps the extent of hybridization of the amplified products on two classes of oligonucleotides is determined by detection of the label of the amplified products, and a conclusion is made on the extent of methylation of said specific cytosine in the genomic DNA sample from the ratio of the labels detected on the two classes of oligonucleotides as a consequence of the hybridization.

Abstract: A method is described for the detection of the degree of methylation of a specific cytosine in the sequence context 5?-CpG-3? of a genomic DNA sample. In the first step, the genomic DNA is chemically treated in such a way that the cytosine bases are converted to uracil, but not the 5-methylcytosine bases. Then segments of the genomic DNA which contain the said specific cytosine are amplified, whereby the amplified products are given a detectable label and in the following steps the extent of hybridization of the amplified products on two classes of oligonucleotides is determined by detection of the label of the amplified products, and a conclusion is made on the extent of methylation of said specific cytosine in the genomic DNA sample from the ratio of the labels detected on the two classes of oligonucleotides as a consequence of the hybridization.

Abstract: The present invention relates to a method for the analysis of methylated cytosines in DNA. In the first step of the invention unmethylated cytosines in the DNA to be analysed are chemically converted into uracil while 5-methylcytosines remain unchanged. In a second step a methylation specific oligonucleotide carrying a non-extendable 3? end is annealed to the converted DNA. Subsequently, the non-extendable 3? terminus of the oligonucleotide is removed in case the oligonucleotide is bound to the DNA with the methylation status to be detected. Finally the unblocked oligonucleotide is extended, and the methylation status is concluded from the absence or presence of an extended oligonucleotide product. The method is preferably used for diagnosis and/or prognosis of adverse events for individuals, for distinguishing cell types and tissues, or for investigating cell differentiation.

Abstract: Particular aspects of the present invention provide a method for quantification of two different variations of a DNA sequence. Particularly, the invention relates to a quantification of methylated DNA, and for this purpose, the test DNA is converted so that cytosine is converted to uracil, while 5-methylcytosine remains unchanged. The converted DNA is amplified by means of a real-time PCR, wherein two labeled real-time probe types are utilized: one specific for methylated DNA; and one for unmethylated DNA. Preferably, the degree of methylation of the test DNA is calculated from the ratio of the signal intensities of the probes or from the Ct values. The inventive methods have substantial utility for diagnosis and prognosis of cancer and other disorders associated with altered or characteristic DNA methylation status, as well as having substantial utility for analysis of SNPs, allelic expression, and prediction of drug response, drug interactions, among other uses.

Abstract: The invention relates to a method for detecting specific dinucleotides in a DNA-sample. A polymerase-chain reaction (PCR) is carried out by using a) a nucleotide, which is part of the dinucleotide which is to be detected, wherein an adequate quantity thereof is marked by a donor-fluorphore and b) another nucleotide, which is part of the dinucleotide which is to be detected, wherein an adequate quantity thereof is marked with an acceptor-fluorophore. Said method determines or quantifies the presence of the dinucleotide by measuring the dimensions of the fluorescence resonance energy transfer (FRET) between the donor- and acceptor-fluorophore.