Patent · US Active

5-methylcytosine detection, compositions and methods therefor

US7999091B2 · kind B2 · utility

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Key dates

Filing dateJun 10, 2008
Grant dateAug 16, 2011
Priority date
Expiry dateJun 10, 2028

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC12Q1/6869
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

Compositions and methods for detecting 5-methylcytosine in a nucleic acid are disclosed. A 5-methylcytosine discriminator, which is a deoxyribonucleosidetriphosphate comprising a cytosine-pairing moiety such as a guanosine and a moiety which hinders hydrogen bonding between the cytosine-pairing moiety and a 5-methylcytosine is described. The discriminator is able to base pair with a cytosine but not a 5-methylcytosine. A 5-methylcytosine comprised by a target nucleotide can be detected in a reaction using a DNA polymerase and a primer hybridized immediately adjacent to the target nucleotide. In the reaction, pyrophosphate released upon incorporation of a dNTP complementary to a target nucleotide is detected. Lack of incorporation of the discriminator, but incorporation of a dGTP, can indicate that the target nucleotide is a 5-methylcytosine.

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