integrated purification and measurement of DNA methylation and commensuration of mrna mutations and / or expression levels in an automated reaction cartridge

摘要

Methods for determining DNA methylation are provided. in an illustrative, but not limiting embodiment, the method comprises i) contacting a biological sample comprising a nucleic acid into a first matrix material comprising a first column or filter, wherein said matrix material binds and / or filters nucleic acids into said sample, thereby purifying the DNA; ii) eluting the bound DNA from the first matrix material and denaturing the DNA to produce eluted denatured DNA; iii) heating the eluted DNA in the presence of bisulfite ions to produce a deaminated nucleic acid; iv) contacting said deaminate nucleic acid with a second matrix material comprising a second column for attaching said deaminate nucleic acid to said second matrix material; v) desulfonating the bound deaminated nucleic acid and / or simultaneously eluting and desulphonating the nucleic acid by bringing the deaminated nucleic acid into contact with an alkaline solution to produce a bisulfite converted nucleic acid; vi) eluting said bisulfite converted nucleic acid from said second matrix material.