Compositions for chromosome-specific staining
First Claim
1. Nucleic acid probes for use in in situ hybridization comprising a heterogeneous mixture that contains labeled nucleic acid fragments that are substantially complementary to unique nucleic acid segments, wherein said mixture comprises blocking nucleic acid having nucleic acid fragments which are substantially complementary to repetitive segments in the labeled nucleic acid, produced by the process of:
- (a) obtaining chromosome-specific DNA fragments;
(b) amplifying said chromosome-specific DNA fragments;
(c) labeling said chromosome-specific DNA fragments with a fluorescent or affinity label; and
(d) adding sufficient blocking nucleic acid to prevent substantial binding of the labeled nucleic acid sequences to repetitive nucleic acid sequences in a target DNA.
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Accused Products
Abstract
Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.
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Citations
13 Claims
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1. Nucleic acid probes for use in in situ hybridization comprising a heterogeneous mixture that contains labeled nucleic acid fragments that are substantially complementary to unique nucleic acid segments, wherein said mixture comprises blocking nucleic acid having nucleic acid fragments which are substantially complementary to repetitive segments in the labeled nucleic acid, produced by the process of:
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(a) obtaining chromosome-specific DNA fragments; (b) amplifying said chromosome-specific DNA fragments; (c) labeling said chromosome-specific DNA fragments with a fluorescent or affinity label; and (d) adding sufficient blocking nucleic acid to prevent substantial binding of the labeled nucleic acid sequences to repetitive nucleic acid sequences in a target DNA. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification