COMPOSITION AND METHODS RELATED TO MODIFICATION OF 5-METHYLCYTOSINE (5-mC)
First Claim
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1. A method for detecting 5-methylcytosine in a nucleic acid comprising converting 5-methylcytosine to 5-hydroxymethylcytosine, 5-carboxycytosine, or 5-formylcytosine with a methylcytosine dioxygenase or the catalytic domain of a methylcytosine dioxygenase and sequencing the nucleic acid to detect 5-mC detecting 5-hydroxymethylcytosine.
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Abstract
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.
30 Citations
120 Claims
- 1. A method for detecting 5-methylcytosine in a nucleic acid comprising converting 5-methylcytosine to 5-hydroxymethylcytosine, 5-carboxycytosine, or 5-formylcytosine with a methylcytosine dioxygenase or the catalytic domain of a methylcytosine dioxygenase and sequencing the nucleic acid to detect 5-mC detecting 5-hydroxymethylcytosine.
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80. A method for distinguishing 5-hydroxymethylcytosine (5-hmC) from 5-methylcytosine (5-mC) in a nucleic acid molecule comprising:
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a) modifying 5-hydroxymethylcytosine in the nucleic acid molecule; b) treating the nucleic acid molecule with methylcytosine dioxygenase to oxidize 5-mC; and
, under conditions to convert 5-methylcytosine to 5-carboxylcytosine;c) then sequencing the treated nucleic acid to identify 5-hmC and 5-mC using bisulfite. - View Dependent Claims (82, 85, 86, 91)
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111. A method for detecting 5-methylcytosine (5-mC) in a nucleic acid molecule comprising
converting 5-mC to a modified 5-mC using enzymatic modification by a methylcytosine dioxygenase; - and,
detecting modified 5-mC by sequencing. - View Dependent Claims (112, 113, 114, 115, 116, 117, 118, 119, 120)
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Specification