Improved method for bisulfite treatment

Procede ameliore pour traiter l'hydrogenosulfite

Abstract

The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. rnethylated 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.
L'invention concerne un procédé pour améliorer la réaction bisulfitique, permettant de déterminer les positions de méthylation dans un acide nucléique, en particulier des cytosines méthylées ou non méthylées, ledit acide nucléique étant incubé dans une solution comprenant l'acide nucléique pour une durée comprise entre 1,5 et 3,5 heures à une température comprise entre 70 et 90 °C. La concentration d'hydrogénosulfite dans la solution est comprise entre 3M et 6,25M, la valeur du pH de la solution étant comprise entre 5,0 et 6,0. Ledit acide nucléique, en particulier les bases de cytosine dans l'acide nucléique sont désaminées. Puis, la solution comprenant l'acide nucléique désaminé est désulfatée et de préférence désalée. L'invention concerne également une solution comprenant de l'hydrogénosulfite comportant un certain pH, l'utilisation de ladite solution et un kit comprenant la solution.

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