Detection of nucleic acid sequence differences by comparative genomic hybridization
First Claim
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1. An array Comparative Genomic Hybridization (CGH) method of detecting one or more nucleotide sequence differences in nucleic acid sequences in a first sample relative to nucleic acid sequences in a second sample, said method comprising:
- (a) labeling nucleic acids from each sample with a different label;
(b) contacting the labeled nucleic acids from each sample with target nucleic acids, wherein either the labeled nucleic acids or the target nucleic acids, or both, have had repetitive sequences, if initially present, blocked and/or removed; and
(c) comparing the intensities of the signals from labeled nucleic acids hybridized to the target nucleic acids to detect one or more nucleotide sequence differences between the samples, wherein;
said contacting comprises contacting the labeled nucleic acids from each sample with an array of target elements comprising the target nucleic acids;
said comparing comprises comparing the intensities of the signals from labeled nucleic acids hybridized to each target element; and
the sequence complexity of each target element is greater than about 50 kilobases and the sequence divergence between the samples is less than about 10%.
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Abstract
The present invention provides a method of detecting nucleotide sequence differences between two nucleic acid samples. The method employs a comparative genomic hybridization (CGH) technique to analyze the sequence differences between the samples. This method permits the identification of small sequence differences (e.g., sequence divergence of 1% or less) in nucleic acid samples of high complexity (e.g., an entire genome).
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Citations
33 Claims
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1. An array Comparative Genomic Hybridization (CGH) method of detecting one or more nucleotide sequence differences in nucleic acid sequences in a first sample relative to nucleic acid sequences in a second sample, said method comprising:
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(a) labeling nucleic acids from each sample with a different label; (b) contacting the labeled nucleic acids from each sample with target nucleic acids, wherein either the labeled nucleic acids or the target nucleic acids, or both, have had repetitive sequences, if initially present, blocked and/or removed; and (c) comparing the intensities of the signals from labeled nucleic acids hybridized to the target nucleic acids to detect one or more nucleotide sequence differences between the samples, wherein; said contacting comprises contacting the labeled nucleic acids from each sample with an array of target elements comprising the target nucleic acids; said comparing comprises comparing the intensities of the signals from labeled nucleic acids hybridized to each target element; and the sequence complexity of each target element is greater than about 50 kilobases and the sequence divergence between the samples is less than about 10%. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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Specification