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    • 9. 发明申请
    • COMPOSITION AND METHODS RELATED TO MODIFICATION OF 5-METHYLCYTOSINE (5mC)
    • WO2012138973A3
    • 2012-10-11
    • PCT/US2012/032489
    • 2012-04-06
    • THE UNIVERSITY OF CHICAGOHE, ChuanZHANG, LiangSONG, ChunxiaoYU, Miao
    • HE, ChuanZHANG, LiangSONG, ChunxiaoYU, Miao
    • C12Q1/68
    • The present invention relates to methods and compositions for detecting, evaluating, and/or mapping 5 methyl-modified and/or 5-hydroxylmethyl-modified cytosine bases within a nucleic acid molecule. In certain aspects the methods further comprise modifying 5hmC with a first detectable label or a first detectable functional group. In certain aspects a converted or modified 5-methylcytosine is labeled with a second detectable label. The method can also include a step of detecting differential labeling of the nucleic acid with the first and second label. The label or detectable label can be a fluorescent radioactive, enzymatic, electrochemical, or colorimetric label. In certain aspects the hydroxyl group of a modified 5mC is further converted to a functional group selected from an aldehyde or carboxyl group, which may in turn be coupled to a label or detectable label. The functional group, e.g., hydroxyl group, of modified 5mC can be further modified using an enzymatic method, such as modification by alcohol dehydrogenase. The functional group, e.g., hydroxyl group, of modified 5mC can be further modified using a chemical method, such as modification by pyridinium chlorochromate (PCC). In certain aspects 5hmC, endogenous and/or modified, is glucosylated. The glucosylated 5hmC (5gmC) can comprise a first, second, and or third label, or more. In certain aspects the label is fluorescent, radioactive, enzymatic, electrochemical, or colorimetric.