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US 11,078,529 B2Granted
Methods and kits for detection of methylation status
AssigneeOslo Universitetssykehus Hf
InventorsJohn Arne Dahl, Adam Brian Robertson, Arne Klungland, Linda Ellevog
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Abstract
The present invention relates to methods and kits for the detection of 5-hydroxymethylcytosine (5hmC) and/or 5-methylcytosine (5meC). In some embodiments, the present invention relates to methods and kits for detection of 5hmC and/or 5meC in nucleic acid (e.g., DNA, RNA). In some embodiments, the present invention relates to detection of 5hmC in genomic DNA, e.g., mammalian genomic DNA.
Claims15
A method for detecting 5-methylated cytosine and 5-hydroxymethylated cytosine of a nucleic acid sequence, wherein said method comprising:
- (a) contacting a nucleic acid methyltransferase with said nucleic acid sequence and a daughter sequence of said nucleic acid sequence, which daughter sequence comprises a complementary sequence of said nucleic acid sequence, such that a pattern of 5-methylated cytosine residues of said nucleic acid sequence is maintained;
- (b) after (a), oxidizing unmodified cytosine of said nucleic acid sequence and of said daughter sequence to yield a modified nucleic acid sequence and a modified daughter sequence;
- (c) sequencing said modified nucleic acid sequence or derivative thereof and said modified daughter sequence or derivative thereof to obtain a first sequence corresponding to said modified nucleic acid sequence and a second sequence corresponding to said modified daughter sequence;
- and
- (d) comparing said first sequence and said second sequence to identify and distinguish said 5-methylated cytosine and said 5-hydroxymethylated cytosine of said nucleic acid sequence.
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Dependencies are inferred from the claim text.