http://rdf.ncbi.nlm.nih.gov/pubchem/patent/JP-2004523201-A
Outgoing Links
| Predicate | Object |
|---|---|
| classificationCPCAdditional | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2600-156 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6841 |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6876 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N15-09 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N21-78 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N33-53 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N33-566 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G01N33-58 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-68 |
| filingDate | 2001-05-15^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationDate | 2004-08-05^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | JP-2004523201-A |
| titleOfInvention | Single copy genome hybridization probe and method for producing the same |
| abstract | The present invention solves the above-mentioned problems and provides a nucleic acid (eg, DNA) hybridization probe containing a labeled single-copy nucleic acid, which hybridizes with a sequence region suspected of being a single copy of a known target nucleic acid. Things. In general, the probes of the present invention are designed by comparing the sequence of a target nucleic acid to known repeat sequences in the genome of which the target is a part; a single copy sequence (ie, a specific There is information that can be derived as a sequence that does not contain a repetitive sequence at all because of its lack of homology, which may block the hybridization signal of a single-copy sequence). As you might imagine, this early phase requires knowledge of the target and repetitive sequences on the genome, and this information could be exploded by the human genome project and related bioinformatics studies. It is becoming. In addition, commercially available computer software can be used to search for the required single copy sequence. As used herein, a probe is most preferably complementary to a target sequence; that is, a probe nucleotide and a target sequence have 100% complementarity. More broadly, one that has less than 100% complementarity can be used as long as the probe hybridizes properly with the target sequence; that is, the probe and the sequence that is the complement of the target sequence are: It is desirable that there be at least 80% homology. More preferably, it is at least about 90%. A nucleic acid fragment corresponding to a sequence presumed to be a single copy can be prepared by various methods such as PCR amplification, treatment of a purified genomic fragment with a restriction enzyme or exonuclease, or direct synthesis of a nucleic acid. Thereafter, the single-copy nucleic acid fragments are purified, for example, by electrophoresis or high performance liquid chromatography, to remove potentially contaminating repetitive sequences; this process is due to false positives of irrelevant genomic sequences. It is particularly preferred to do so to eliminate hybridization and detection. The probe fragment can then be cloned into a recombinant DNA vector or directly labeled. Probes are preferably labeled by nick translation using modified or directly labeled nucleotides. Thereafter, the purified nucleic acid, preferably immobilized on a microslide, or otherwise immobilized on a membrane, slide, DNA chip, or other object, and the labeled probe are denatured, Hybridize. Probes hybridize to chromosomes by conventional fluorescent staining in situ hybridization (FISH), as described in US Pat. Nos. 5,985,549 and 5,478,841; By the described technique, it can also be bound to the immobilized nucleic acid. The signal of the probe can be visualized by various methods, such as using fluorescent, immunological, or enzymatic reaction solutions. The use of the probe of the present invention allows the chromosomal breakpoint to be more accurately determined with a resolution that could not be achieved by conventional in situ hybridization. In performing such an analysis, a set of probes to be used initially can be prepared based on the region considered to be on the opposite side of the breakpoint. After the initial experiment to determine the breakpoint, a new probe can be designed near the breakpoint using the single copy approach. In this way, it is possible to determine the exact area of the cutting point. It was revealed that the presence of a repeat sequence in the genome, which was hitherto unknown, can be determined using a probe that is assumed to be a single copy. The previously unknown family of repeating sequences is added to a database of contiguous nucleic acid sequences, which can then be used to design single copy probes. It has been clarified that the probe of the present invention may contain a sequence that has been doubled or triplicated in the genome so that hybridization can be performed more firmly due to the longer target sequence. . Further, such a duplicon or triplicon can be confirmed using, for example, a single copy probe, and it is difficult to use a commercially available probe. |
| isCitedBy | http://rdf.ncbi.nlm.nih.gov/pubchem/patent/JP-2010522571-A http://rdf.ncbi.nlm.nih.gov/pubchem/patent/JP-2008534011-A |
| priorityDate | 2000-05-16^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
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