http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2018046521-A1
Outgoing Links
| Predicate | Object |
|---|---|
| assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_1b5d47c4f1d77d82de7ce1f64ba985b3 |
| classificationCPCAdditional | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2565-1015 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2521-101 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2563-107 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2521-307 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2533-107 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2537-149 |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6823 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6818 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6869 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-68 |
| filingDate | 2017-09-06-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_28f845b0c76a27e4f9f4e2224ffda0cd http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_9fa5e6e5c90e8f185648f8fbe551d30f |
| publicationDate | 2018-03-15-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | WO-2018046521-A1 |
| titleOfInvention | Single nucleotide detection method and associated probes |
| abstract | A method of sequencing a nucleic acid is provided. It is characterised by the steps of (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one substantially double-stranded primary oligonucleotide used probe by reacting, in the presence of a polymerase and a ligase, at least one of the single nucleoside triphosphates with a corresponding primary probe comprising (a) a first single- stranded oligonucleotide including a restriction endonuclease nicking-site, a single nucleotide capture site for capturing the single nucleoside triphosphate and oligonucleotide flanking regions juxtaposed either side of the capture site and (b) second and third single-stranded oligonucleotides capable of hybridising to the first oligonucleotide flanking regions; (3) nicking the first oligonucleotide strand of the used primary probe at the nicking-site with a nicking restriction endonuclease to create separate first oligonucleotide components; (4) separating the first oligonucleotide components generated in step (3) from the complementary strand of the used probe; (5) producing at least one substantially double-stranded secondary used probe by reacting, in the presence of a ligase, at least one of the separated first oligonucleotide components with a corresponding secondary probe comprising (c) a complementary fourth oligonucleotide bearing fluorophores in a substantially undetectable state and optionally (d) a fifth oligonucleotide at least in part complementary to the fourth oligonucleotide; (6) digesting the used secondary probe with an enzyme having double-stranded exonucleolytic activity to yield the fluorophores in a detectable state and a single-stranded sixth oligonucleotide which is at least in part the sequence complement of the fourth oligonucleotide and (7) detecting the fluorophores released in step (6). The method is advantageously carried out in microdroplets. Corresponding biological probe systems comprised of the primary and secondary probes are also described. |
| isCitedBy | http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-11318472-B2 |
| priorityDate | 2016-09-06-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
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