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AssigneeThe Children's Mercy Hospital
InventorsJoan H. M. Knoll, Peter K. Rogan
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Nucleic acid (e.g., DNA) hybridization probes are described which comprise a labeled, single copy nucleic acid which hybridizes to a deduced single copy sequence interval in target nucleic acid of known sequence. The probes, which are essentially free of repetitive sequences, can be used in hybridization analyses without adding repetitive sequence-blocking nucleic acids. This allows rapid and accurate detection of chromosomal abnormalities. The probes are preferably designed by first determining the sequence of at least one single copy interval in a target nucleic acid sequence, and developing corresponding hybridization probes which hybridize to at least a part of the deduced single copy sequence. In practice, the sequences of the target and of known genomic repetitive sequence representatives are compared in order to deduce locations of the single copy sequence intervals. The single copy probes can be developed by any variety of methods, such as PCR amplification, restriction or exonuclease digestion of purified genomic fragments, or direct synthesis of DNA sequences. This is followed by labeling of the probes and hybridization to a target sequence.
A nucleic acid hybridization probe comprising a labeled, single copy nucleic acid which will hybridize to a deduced single copy sequence interval in target nucleic acid of known sequence, said nucleic acid probe having a length of at least about 50 nucleotides.
In a hybridization method including the steps of preparing a reaction mixture comprising a target nucleic acid sequence and a nucleic acid probe which hybridizes to at least a portion of said target nucleic acid sequence, and causing said probe to hybridize to said target nucleic acid sequence, the improvement which comprises using as said probe a labeled, single copy nucleic acid which hybridizes to a deduced single copy sequence interval in target nucleic acid of known sequence, said nucleic acid probe having a length of at least about 50 nucleotides.
A method of developing a hybridization probe for a target nucleic acid sequence forming a part of a genome, said method comprising the steps of:
A DNA sequence having at least about 80% sequence identity with a member selected from the group consisting of SEQ ID Nos. 429-446.
A DNA sequence selected from the group consisting of SEQ ID Nos. 429-446 and 480-613.
A DNA sequence having at least about 80% sequence identity with a PCR-amplified sequence produced using as PCR primers adjacent pairs of the sequences identified as SEQ ID Nos. 429-446 and 480-613, beginning with SEQ ID No. 429.
A method of determining the existence of previously unknown repeat sequence families in a genome, comprising the steps of reacting a labeled, putative single copy nucleic acid probe with the genome, causing the probe to hybridize, and ascertaining if the probe hybridizes to the genome at more than three different locations as a determination of a new repeat sequence family.
A method of determining a chromosome breakpoint, comprising the steps of:
Dependencies are inferred from the claim text.