Methods for detection of nucleotide modification

US10428381B2 · kind B2 · utility

Assignee

• Cambridge Epigenetix Limited · GB

Inventors

• Michael John Booth · Derby, GB
• Shankar Balasubramanian · Cambridge, GB

Key dates

• Filing date: Mar 21, 2016
• Grant date: Oct 1, 2019
• Expiry date: Feb 24, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/154
• WIPO field: Organic fine chemistry
• WIPO sector: Chemistry

Abstract

This invention relates to the identification of modified cytosine residues, such as 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC) and 5-formylcytosine (5fC) to be distinguished from cytosine (C) in a sample nucleotide sequence. Methods may comprise oxidizing or reducing a first portion of polynucleotides which comprise the sample nucleotide sequence; treating the oxidized or reduced first portion and a second portion of polynucleotides with bisulfite; sequencing the polynucleotides in the first and second portions of the population following steps ii) and iii) to produce first and second nucleotide sequences, respectively and; identifying the residue in the first and second nucleotide sequences which corresponds to a cytosine residue in the sample nucleotide sequence. These methods may be useful, for example in the analysis of genomic DNA and/or of RNA.