Method for bisulfite treatment

US7413855B2 · kind B2 · utility

Assignee

• Roche Molecular Systems, Inc. · US

Inventors

• Frank Bergmann · Iffeldorf, DE
• Christine Markert-Hahn · Penzberg, DE
• Joerg Kleibler · Penzberg, DE
• Dirk Block · Bichl, DE

Key dates

• Filing date: Jan 28, 2004
• Grant date: Aug 19, 2008
• Expiry date: Jan 28, 2024

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6827
• WIPO field: Organic fine chemistry
• WIPO sector: Chemistry

Abstract

The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. methylated and non-methylated cytosines, whereby the nucleic acid is incubated in a solution comprising the nucleic acid for a time period of 1.5 to 3.5 hours at a temperature between 70 and 90° C., whereby the concentration of bisulfite in the solution is between 3 M and 6.25 M and whereby the pH value of the solution is between 5.0 and 6.0 whereby the nucleic acid, i.e. the cytosine bases in the nucleic acid, are deaminated. Then the solution comprising the deaminated nucleic acid is desulfonated and preferably desalted. The application is further related to a solution comprising bisulfite with a certain pH and uses thereof as well as a kit comprising the solution.