Patent · US Expired

5-methylcytosine detection, compositions and methods therefor

US7399614B2 · kind B2 · utility

38Cited by
5References
29Claims
0Family size

Assignee

Inventor

Key dates

Filing dateJun 30, 2004
Grant dateJul 15, 2008
Priority date
Expiry dateJan 19, 2025

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.

Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.