http://rdf.ncbi.nlm.nih.gov/pubchem/patent/EP-3211092-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/C12Q2521-331 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2521-319 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2521-501 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2525-301 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2563-159 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2565-629 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2525-131 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2565-107 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2525-125 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2525-307 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q2565-301 |
| classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6827 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12Q1-6869 |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12Q1-68 |
| filingDate | 2016-02-24-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_9fa5e6e5c90e8f185648f8fbe551d30f |
| publicationDate | 2017-08-30-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | EP-3211092-A1 |
| titleOfInvention | Single nucleotide detection method |
| abstract | A method of sequencing a nucleic acid characterised by the steps of (1) generating a stream of single nucleotides by progressive pyrophosphorolysis of the nucleic acid; (2) producing at least one substantially double-stranded oligonucleotide used probe by reacting, in the presence of a polymerase and a ligase, one of the single nucleotides with a corresponding probe system comprising (a) a first single-stranded oligonucleotide labelled with first and second regions of characteristic detectable element types in an undetectable state located respectively on the X' and Y' end sides of a third region comprising a restriction enzyme recognition site element including the capture site and an exonuclease-blocking site on the X' side thereof (wherein either X' is 3' and Y' is 5' or X' is 5' and Y' is 3') and (b) second and third single-stranded oligonucleotides capable of hybridising to complementary regions on the first oligonucleotide flanking the capture site; (2a) either (i) treating the used probe with a conventional or nicking substitution-dependent restriction endonuclease to cut the first oligonucleotide strand at the recognition site if and only if the single nucleotide captured comprises a nucleobase which is substituted or (ii) treating the used probe with a conventional or nicking substitution-sensitive restriction endonuclease to cut the first oligonucleotide strand at the recognition site if and only if the single nucleotide captured comprises a nucleobase which is unsubstituted; (3) digesting the first oligonucleotide strand of the used probe with an enzyme having double-stranded exonucleolytic activity in the X'-Y' direction corresponding to the first oligonucleotide to yield detectable elements derived from either the first region, the second region, or the first and second regions in a detectable state and a single-stranded fourth oligonucleotide which is at least in part the sequence complement of the first oligonucleotide; (4) reacting the fourth oligonucleotide with another first oligonucleotide to produce a substantially double-stranded oligonucleotide product corresponding to the used probe; (5) repeating steps (2a), (3) and (4) in a cycle and (6) detecting the detectable elements released in each iteration of step (3) wherein if the endonuclease employed is of the conventional type the second or third oligonucleotide includes an endonucleolysis directing linkage at or close to its X' or Y' end respectively. n Corresponding biological probe systems are also disclosed. |
| isCitedBy | http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-10961569-B2 http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-11634754-B2 http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-11332780-B2 |
| priorityDate | 2016-02-24-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
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