Method for bisulfite treatment

摘要

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.

主权项

1. A method for the conversion of cytosine bases in a DNA molecule to uracil bases, comprising:
a) binding a double stranded DNA directly to an unmodified solid phase without a compound mediating the binding to the solid phase, wherein the unmodified solid phase is selected from the group consisting of glass, silica, and a glass or silica containing material, b) denaturing the double stranded DNA, c) incubating the solid phase bound DNA in the presence of sulfite ions whereby the DNA is deaminated, yielding a solid phase bound deaminated DNA, d) optionally washing the solid phase bound deaminated DNA, e) incubating the solid phase bound deaminated DNA under alkaline conditions whereby the deaminated DNA is desulfonated, f) washing the solid phase bound deaminated and desulfonated DNA, and g) eluting the washed deaminated and desulfonated DNA from the solid phase.