http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2018042028-A1

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filingDate 2017-09-04-04:00^^<http://www.w3.org/2001/XMLSchema#date>
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publicationDate 2018-03-08-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber WO-2018042028-A1
titleOfInvention Single nucleotide detection method and associated probes
abstract A method of sequencing a nucleic acid such as DNA or RNA 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 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 biological 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 regions juxtaposed either side of the nicking-site bearing respectively at least one fluorophore and at least one quencher so as to render the fluorophores quenched and (b) second and third single-stranded oligonucleotides capable of hybridising to complementary regions on the first oligonucleotide either side of the capture site; (3) nicking the first oligonucleotide strand of the used probe at the nicking-site with a nicking restriction endonuclease to create separate first oligonucleotide components respectively bearing the fluorophores and the quenchers; (4) separating the first oligonucleotide components generated in step (3) from the complementary strand of the used probe and (5) detecting the fluorophores on the separated oligonucleotide component bearing them. The method is suitable for being performed in microdroplets. New biological probes and probe systems for use with the method are also described.
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http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP65748
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http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP65751
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Total number of triples: 420.