Method for detecting cytosine methylation in DNA samples

US7524629B2 · kind B2 · utility

Assignee

• EPIGENOMICS AG · DE

Inventors

• Alexander Olek · Berlin, DE
• Kurt Berlin · Stahnsdorf, DE

Key dates

• Filing date: Feb 23, 2001
• Grant date: Apr 28, 2009
• Expiry date: Aug 15, 2021

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6858
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Described is a method for detecting 5-methylcytosine in genomic DNA samples. First, a genomic DNA from a DNA sample is chemically converted with a reagent, 5-methylcytosine and cytosine reacting differently, and the pretreated DNA is subsequently amplified using a polymerase and at least one primer. In the next step, the amplified genomic DNA is hybridized to at least one oligonucleotide, forming a duplex, and said oligonucleotide is elongated by at least one nucleotide, the nucleotide carrying a detectable label, and the elongation depending on the methylation status of the specific cytosine in the genomic DNA sample. In the next step, the elongated oligonucleotides are analyzed for the presence of the label.