Differential enzymatic fragmentation by whole genome amplification
First Claim
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1. A method of amplifying unmethylated or methylated DNA fragments in a biological sample, the method comprising:
- (a) providing randomly fragmented DNA from the biological sample;
(b) adding a sequence tag onto at least one end of the DNA fragments, thereby generating modified DNA fragments;
(c) digesting the DNA fragments with a methylation-dependent restriction enzyme or a methylation-sensitive restriction enzyme to obtain intact DNA fragments and digested DNA fragments; and
(d) after the digesting step, amplifying the intact modified DNA fragments with at least one primer that initiates amplification from the sequence tags, thereby generating amplified intact DNA fragments representing the unmethylated or methylated modified intact DNA fragments in the sample.
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Abstract
The present invention provides methods for detecting the presence of methylation at a locus within a population of nucleic acids.
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Citations
20 Claims
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1. A method of amplifying unmethylated or methylated DNA fragments in a biological sample, the method comprising:
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(a) providing randomly fragmented DNA from the biological sample; (b) adding a sequence tag onto at least one end of the DNA fragments, thereby generating modified DNA fragments; (c) digesting the DNA fragments with a methylation-dependent restriction enzyme or a methylation-sensitive restriction enzyme to obtain intact DNA fragments and digested DNA fragments; and (d) after the digesting step, amplifying the intact modified DNA fragments with at least one primer that initiates amplification from the sequence tags, thereby generating amplified intact DNA fragments representing the unmethylated or methylated modified intact DNA fragments in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification