Method for detecting cytosine methylations
AssigneeEpigenomics AG
InventorKurt Berlin
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Abstract
A method is described for the detection of 5-methylcytosine in genomic DNA samples. First, a genomic DNA from a DNA sample is chemically converted with a reagent, whereby 5-methylcytosine and cytosine react differently. Then the pretreated DNA is amplified with the use of a polymerase with primers of different sequence. In the next step, the amplified genomic DNA is hybridized to an oligonucleotide array and PCR products are obtained, which must be provided with a label. Alternatively, the PCR products can be extended in a primer extension reaction, wherein the extension products are also provided with a label. In the last step, the extended oligonucleotides are investigated for the presence of the label.
Claims24
A method for the detection of cytosine methylations in DNA is hereby characterized in that the following operating steps are conducted:
- a) a genomic DNA sample is incubated with a solution of a bisulfite in the concentration range between 0.1 and 6 mol/liter in the presence of at least one radical trap and at least one denaturing solvent;
- b) the treated DNA sample is diluted with water or an aqueous solution;
- c) the DNA sample is amplified in a polymerase reaction to yield amplified DNA;
- d) one detects how much the sequence has changed by the treatment according to step a) in comparison to the genomic DNA sample and concludes the methylation state of at least one locus in the genomic DNA sample.
Dependencies are inferred from the claim text.