http://rdf.ncbi.nlm.nih.gov/pubchem/patent/NZ-604338-A

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filingDate 2012-01-11-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_34ac7f5c13a90df324db97c714067048
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publicationDate 2015-05-29-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber NZ-604338-A
titleOfInvention Detection of target nucleic acid sequences by pto cleavage and extension assay
abstract Disclosed is a method for detecting a target nucleic acid sequence from a DNA or a mixture of nucleic acids by a PTOCE (PTO Cleavage and Extension) assay, comprising: (a) hybridising the target nucleic acid sequence with an upstream oligonucleotide and a PTO (Probing and Tagging Oligonucleotide); wherein the upstream oligonucleotide comprises a hybridising nucleotide sequence complementary to the target nucleic acid sequence; the PTO comprises (i) a 3’-targeting portion comprising a hybridising nucleotide sequence complementary to the target nucleic acid sequence and (ii) a 5’-tagging portion comprising a nucleotide sequence non-complementary to the target nucleic acid sequence; wherein the 3’-targeting portion is hybridised with the target nucleic acid sequence and the 5’-tagging portion is not hybridised with the target nucleic acid sequence; the upstream oligonucleotide is located upstream of the PTO; (b) contacting the resultant of the step (a) to an enzyme having a 5’ nuclease activity under conditions for cleavage of the PTO; wherein the upstream oligonucleotide or its extended strand induces cleavage of the PTO by the enzyme having the 5’ nuclease activity such that the cleavage releases a fragment comprising the 5’-tagging portion or a part of the 5’-tagging portion of the PTO; (c) hybridising the fragment released from the PTO with a CTO (Capturing and Templating Oligonucleotide); wherein the CTO comprises in a 3’ to 5’ direction (i) a capturing portion comprising a nucleotide sequence complementary to the 5’- tagging portion or a part of the 5’-tagging portion of the PTO and (ii) a templating portion comprising a nucleotide sequence non-complementary to the 5’-tagging portion and the 3’-targeting portion of the PTO; wherein the fragment released from the PTO is hybridised with the capturing portion of the CTO; (d) performing an extension reaction using the resultant of the step (c) and a template-dependent nucleic acid polymerase; wherein the fragment hybridised with the capturing portion of the CTO is extended and an extended duplex is formed; wherein the extended duplex has a Tm value adjustable by (i) a sequence and/or length of the fragment, (ii) a sequence and/or length of the CTO or (iii) the sequence and/or length of the fragment and the sequence and/or length of the CTO; (e) melting the extended duplex over a range of temperatures to give a target signal indicative of the presence of the extended duplex; wherein the target signal is provided by (i) at least one label linked to the fragment and/or the CTO, (ii) a label incorporated into the extended duplex during the extension reaction, (iii) a label incorporated into the extended duplex during the extension reaction and a label linked to the fragment and/or the CTO, or (iv) an intercalating label; and (f) detecting the extended duplex by measuring the target signal; whereby the presence of the extended duplex indicates the presence of the target nucleic acid sequence
priorityDate 2011-01-11-04:00^^<http://www.w3.org/2001/XMLSchema#date>
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http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID936617
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB7N8Q1
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID54076
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB2VHN8
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID57569899
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID2320
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID11309
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID88190
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ0TKT1
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP56105
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID88189
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP23989
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID31772786
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP50388
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID855936
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID65134
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA8GGN3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCO32801
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA1JTC4
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ47077
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID469800
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ1PS67
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCO70372
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP45582
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID1261044
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP95690
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ9HT80
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ8LKW0
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID274
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID94694
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419581006
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ47526
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCO33815
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID13945
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID53425817
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID485
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ1KVQ9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ8D4H3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP49676
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2725057
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID94694
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID451065782
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID927419
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID2309
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419525440
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2309
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID1255525
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID2421
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP0A551
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP28040
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP43741
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419563002
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID3169068
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID29292
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2421
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID44129042
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB5Z2P7
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP35396
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ6LL68
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB4S2K9
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID52022
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2287
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ669R9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP23071
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ46666
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA7FH78
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID2287
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP23070
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID856655
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ38087
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID13773
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID836845
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID29292
http://rdf.ncbi.nlm.nih.gov/pubchem/taxonomy/TAXID52023
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419517649
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID24794037
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP23072
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2336
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID2283
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID52023
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCO67779
http://rdf.ncbi.nlm.nih.gov/pubchem/anatomy/ANATOMYID52022
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP19593
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ03181
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID414028668
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID452705

Total number of triples: 375.