Method for relative quantification of methylation of cytosine bases in DNA samples
AssigneeEpigenomics AG
InventorKurt Berlin
Life of this patent
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Abstract
A method is described for the relative quantification of the methylation of cytosine bases in DNA samples, wherein the following method steps are conducted: a) a genomic DNA sample is chemically converted with a reagent, wherein 5-methylcytosine and cytosine react differently and show a different base pairing behavior in the DNA duplex after the reaction; b) the DNA sample is amplified, whereby a fluorescently labeled dCTP or dGTP derivative is added; c) the amplified products are separated spatially from each other; and d) the fluorescence of the separated amplified products is measured quantitatively.
Claims13
A method for the relative quantification of the methylation of cytosine bases in DNA samples, said method comprising the steps of:
- a) chemically reacting a genomic DNA sample with a reagent, wherein 5-methylcytosine and cytosine react differently and these thus show a different base pairing behavior in the DNA duplex after the reaction;
- b) then, amplifying the chemically reacted DNA sample by PCR, said amplifying step comprising the use of a fluorescently labeled dCTP or dGTP derivative to yield amplified products, whereby several fluorophores are introduced into one amplified product;
- c) then, spatially separating the amplified products from each other;
- d) then, quantitatively measuring the fluorescence of the separated amplified products;
- and
- e) then, determining from the measured fluorescence the relative number of methylated cytosine bases that were present in the DNA sample prior to step a).
A method for the relative quantification of the methylation of cytosine bases in DNA samples, said method consisting of the steps of:
- a) chemically reacting a genomic DNA sample with a reagent, wherein 5-methylcytosine and cytosine react differently and these thus show a different base pairing behavior in the DNA duplex after the reaction;
- b) then, amplifying the chemically reacted DNA sample, wherein said amplifying step comprises the use of a fluorescently labeled dCTP or dGTP derivative to yield amplified products;
- c) then, spatially separating the amplified products from each other;
- d) then, quantitatively measuring the fluorescence of the separated amplified products;
- and
- e) then, determining from the measured fluorescence the relative number of methylated cytosine bases that were present in the DNA sample prior to step a).
Dependencies are inferred from the claim text.