Ligase/polymerase method for detecting cytosine methylation in DNA samples

US7405040B2 · 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: Jul 29, 2008
• Expiry date: Apr 29, 2022

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/158
• 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 two oligonucleotides, the latter being joined by inserting at least one oligonucleotide. In the ligation product, one nucleotide carries a detectable label, and the elongation depends 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.