Method for bisulfite treatment
AssigneeEpigenomics AG
InventorsChristine Markert-Hahn, Dirk Block
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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 bound to a solid phase during the deamination and/or desulfonation step of the bisulfite reaction. The solid phase is preferably a material comprising glass or silica, more preferably a glass fleece, glass membrane or a magnetic glass particle. Further, the use of a solid phase for binding a nucleic acid during the deamination and/or desulfonation step of the bisulfite reaction is disclosed and a kit containing a bisulfite reagent and a solid phase.
Claims11
A method for conversion of a cytosine base in a nucleic acid to a uracil base, comprising the steps of:
- a. incubating the nucleic acid in the presence of sulfite ions whereby the nucleic acid is deaminated;
- b. binding the deaminated nucleic acid to a solid phase;
- c. optionally washing the deaminated solid phase bound nucleic acid;
- d. incubating the deaminated solid phase bound nucleic acid under alkaline conditions whereby the deaminated nucleic acid is desulfonated, thereby converting the cytosine base in the nucleic acid to the uracil base;
- e. washing the deaminated and desulfonated solid phase bound nucleic acid;
- and
- f. eluting the washed deaminated and desulfonated nucleic acid from the solid phase.
Dependencies are inferred from the claim text.