Whole genome methylation profiles via single molecule analysis
First Claim
Patent Images
1. A method for identifying a methylation profile in an immobilized, elongated polynucleotide, the method comprising the steps of:
- cleaving the polynucleotide with a methylation-insensitive restriction enzyme;
imaging the cleaved polynucleotide;
further cleaving the imaged polynucleotide with a methylation-sensitive restriction enzyme;
imaging the further cleaved polynucleotide; and
comparing the images and intramolecular position to reference data to identify the methylation profile in the polynucleotide.
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Abstract
Methods for determining methylation profiles of single polynucleotide molecules, especially genomic DNA molecules, by using sequence specificity and methylation sensitivity of restriction endonucleases and other proteins that selectively bind to methylated or unmethylated residues.
42 Citations
10 Claims
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1. A method for identifying a methylation profile in an immobilized, elongated polynucleotide, the method comprising the steps of:
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cleaving the polynucleotide with a methylation-insensitive restriction enzyme;
imaging the cleaved polynucleotide;
further cleaving the imaged polynucleotide with a methylation-sensitive restriction enzyme;
imaging the further cleaved polynucleotide; and
comparing the images and intramolecular position to reference data to identify the methylation profile in the polynucleotide. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for identifying a methylation profile in an immobilized, elongated polynucleotide, the method comprising the steps of:
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indexing the polynucleotide with respect to at least one restriction enzyme cleavage sequence;
selectively binding a labeled methylation-specific reagent to a site on the indexed polynucleotide; and
mapping the labeled reagent to the polynucleotide. - View Dependent Claims (8, 9, 10)
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Specification