AU2002361946A8 - Oligonucleotides comprising intercalator pseudonucleotide(s) for detection of nucleic acids and mutants hereof - Google Patents

Oligonucleotides comprising intercalator pseudonucleotide(s) for detection of nucleic acids and mutants hereof

Info

Publication number
AU2002361946A8
AU2002361946A8 AU2002361946A AU2002361946A AU2002361946A8 AU 2002361946 A8 AU2002361946 A8 AU 2002361946A8 AU 2002361946 A AU2002361946 A AU 2002361946A AU 2002361946 A AU2002361946 A AU 2002361946A AU 2002361946 A8 AU2002361946 A8 AU 2002361946A8
Authority
AU
Australia
Prior art keywords
oligonucleotides
detection
nucleic acids
intercalator pseudonucleotide
mutants hereof
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
AU2002361946A
Other versions
AU2002361946A1 (en
Inventor
Ulf Bech Christensen
Erik Bjerregaard Pedersen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Human Genetic Signatures Pty Ltd
Original Assignee
Human Genetic Signatures Pty Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Human Genetic Signatures Pty Ltd filed Critical Human Genetic Signatures Pty Ltd
Publication of AU2002361946A8 publication Critical patent/AU2002361946A8/en
Publication of AU2002361946A1 publication Critical patent/AU2002361946A1/en
Abandoned legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6832—Enhancement of hybridisation reaction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/13—Decoys
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/318—Chemical structure of the backbone where the PO2 is completely replaced, e.g. MMI or formacetal
    • C12N2310/3181—Peptide nucleic acid, PNA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/321—2'-O-R Modification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/323—Chemical structure of the sugar modified ring structure
    • C12N2310/3231—Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C12N2310/3511—Conjugate intercalating or cleaving agent
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C12N2310/3517—Marker; Tag
AU2002361946A 2001-12-18 2002-12-18 Oligonucleotides comprising intercalator pseudonucleotide(s) for detection of nucleic acids and mutants hereof Abandoned AU2002361946A1 (en)

Applications Claiming Priority (19)

Application Number Priority Date Filing Date Title
DKPA200101897 2001-12-18
DKPA200101900 2001-12-18
DKPA200101898 2001-12-18
DKPA200101897 2001-12-18
DKPA200101898 2001-12-18
DKPA200101899 2001-12-18
DKPA200101899 2001-12-18
DKPA200101900 2001-12-18
US36554502P 2002-03-20 2002-03-20
US60/365,545 2002-03-20
DKPA200201578 2002-10-14
DKPA200201577 2002-10-14
DKPA200201576 2002-10-14
DKPA200201575 2002-10-14
DKPA200201577 2002-10-14
DKPA200201578 2002-10-14
DKPA200201575 2002-10-14
DKPA200201576 2002-10-14
PCT/DK2002/000877 WO2003052134A2 (en) 2001-12-18 2002-12-18 Oligonucleotides comprising intercalator pseudonucleotide(s) for detection of nucleic acids and mutants hereof

Publications (2)

Publication Number Publication Date
AU2002361946A8 true AU2002361946A8 (en) 2003-06-30
AU2002361946A1 AU2002361946A1 (en) 2003-06-30

Family

ID=27576036

Family Applications (4)

Application Number Title Priority Date Filing Date
AU2002358464A Ceased AU2002358464B2 (en) 2001-12-18 2002-12-18 Pseudonucleotide comprising an intercalator
AU2002358463A Abandoned AU2002358463A1 (en) 2001-12-18 2002-12-18 Oligonucleotide analogues comprising intercalator pseudonucleotides
AU2002358462A Abandoned AU2002358462A1 (en) 2001-12-18 2002-12-18 Linear and hairpin oligonucleotide analogues comprising intercalator pseudonucleotides
AU2002361946A Abandoned AU2002361946A1 (en) 2001-12-18 2002-12-18 Oligonucleotides comprising intercalator pseudonucleotide(s) for detection of nucleic acids and mutants hereof

Family Applications Before (3)

Application Number Title Priority Date Filing Date
AU2002358464A Ceased AU2002358464B2 (en) 2001-12-18 2002-12-18 Pseudonucleotide comprising an intercalator
AU2002358463A Abandoned AU2002358463A1 (en) 2001-12-18 2002-12-18 Oligonucleotide analogues comprising intercalator pseudonucleotides
AU2002358462A Abandoned AU2002358462A1 (en) 2001-12-18 2002-12-18 Linear and hairpin oligonucleotide analogues comprising intercalator pseudonucleotides

Country Status (4)

Country Link
EP (1) EP1468007A2 (en)
CN (1) CN1653079B (en)
AU (4) AU2002358464B2 (en)
WO (4) WO2003052133A2 (en)

Families Citing this family (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2003900368A0 (en) * 2003-01-24 2003-02-13 Human Genetic Signatures Pty Ltd Assay for nucleic acid molecules
WO2004074503A2 (en) * 2003-02-21 2004-09-02 Hoser Mark J Nucleic acid sequencing methods, kits and reagents
WO2004101816A1 (en) * 2003-05-15 2004-11-25 Human Genetic Signatures Pty Ltd Decoys for nucleic acid-binding proteins
WO2004111266A1 (en) 2003-06-17 2004-12-23 Human Genetic Signatures Pty Ltd Methods for genome amplification
CA2537810C (en) 2003-09-04 2012-12-18 Human Genetic Signatures Pty Ltd Nucleic acid detection assay
JP4397659B2 (en) * 2003-09-11 2010-01-13 独立行政法人科学技術振興機構 DNA detection method using molecular beacon using switching between monomer emission and excimer emission of fluorescent molecules
US8168777B2 (en) 2004-04-29 2012-05-01 Human Genetic Signatures Pty. Ltd. Bisulphite reagent treatment of nucleic acid
ATE476439T1 (en) * 2004-09-10 2010-08-15 Human Genetic Signatures Pty AMPLIFICATION BLOCKERS COMPRISING INTERCALATING NUCLEIC ACIDS (INA) CONTAINING INTERCALATING PSEUDONUCLEOTIDES (IPN)
EP1799853A2 (en) 2004-09-24 2007-06-27 Universität Bern Molecular beacons
EP1828411B1 (en) 2004-12-03 2012-11-07 Human Genetic Signatures PTY Ltd Methods for simplifying microbial nucleic acids by chemical modification of cytosines
ES2436760T3 (en) * 2005-05-25 2014-01-07 Tina Holding Aps Stable and selective formation of triple helices and double Hoogsteen type propellers using torsion intercalating nucleic acids (TINA) and procedure for the preparation of TINA
JP2008541705A (en) 2005-05-26 2008-11-27 ヒューマン ジェネティック シグネチャーズ ピーティーワイ リミテッド Isothermal strand displacement amplification using primers containing non-standard bases
ATE531820T1 (en) 2005-09-14 2011-11-15 Human Genetic Signatures Pty HEALTH CONDITION TEST
WO2007104318A2 (en) * 2006-03-16 2007-09-20 Pentabase Aps Oligonucleotides comprising signalling pairs and hydrophobic nucleotides, stemless beacons, for detection of nucleic acids, methylation status and mutants of nucleic acids
EP1845166A1 (en) * 2006-04-13 2007-10-17 BioSpring GmbH Nucleic acid probes with covalently bound intercalating fluorescent dyes for hybridization with target nucleic acids
WO2009027978A1 (en) 2007-08-30 2009-03-05 Hadasit Medical Research Services & Development Ltd. NUCLEIC ACID SEQUENCES COMPRISING NF-ϰB BINDING SITE WITHIN O(6)-METHYLGUANINE-DNA-METHYLTRANSFERASE (MGMT) PROMOTER REGION AND USES THEREOF FOR THE TREATMENT OF CANCER AND IMMUNE-RELATED DISORDERS
EP2215250B1 (en) 2007-11-27 2013-02-27 Human Genetic Signatures Pty Ltd Enzymes for amplification and copying bisulphite modified nucleic acids
EP2229457A4 (en) 2007-12-14 2011-01-26 Minitube America Inc Gender-specific separation of sperm cells and embryos
WO2011032034A2 (en) 2009-09-10 2011-03-17 University Of Idaho Nucleobase-functionalized conformationally restricted nucleotides and oligonucleotides for targeting nucleic acids
CN102933720B (en) * 2010-05-07 2015-03-18 昆特拜克股份公司 Method for generating a double stranded nucleic acid with a single stranded overhang
RU2014106024A (en) 2011-07-19 2015-08-27 Юниверсити Оф Айдахо OPTIONS FOR IMPLEMENTING THE PROBE AND METHODS OF DIRECTED ACTION FOR NUCLEIC ACIDS
NZ622418A (en) 2011-09-07 2014-12-24 Human Genetic Signatures Pty Molecular detection assay
DK3350347T3 (en) 2015-09-14 2021-04-19 Pentabase Aps Methods and materials for detection of mutations
WO2018013999A1 (en) 2016-07-15 2018-01-18 Am Chemicals Llc Non-nucleosidic solid supports and phosphoramidite building blocks for oligonucleotide synthesis
AU2019282789A1 (en) * 2018-06-08 2020-12-17 Carnegie Mellon University Modified nucleobases with uniform H-bonding interactions, homo- and hetero-basepair bias, and mismatch discrimination
CN112601825B (en) * 2018-07-09 2023-11-28 青岛华大智造科技有限责任公司 Nucleic acid sequencing method
CN114286865A (en) 2019-05-13 2022-04-05 潘塔贝斯公司 Melting temperature methods, kits and reporter oligonucleotides for detecting variant nucleic acids
CN111073884A (en) * 2020-02-14 2020-04-28 昆明理工大学 Method for improving accuracy of SNP locus detection with functional effect in SNP with distance of non-coding region less than 50bp
WO2022002958A1 (en) 2020-06-29 2022-01-06 Pentabase Aps Detection of methylation status

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3293820B2 (en) * 1985-12-13 2002-06-17 エンゾ− バイオケム インコ−ポレイテツド Novel one-step method and polynucleotide compound for hybridizing to target polynucleotide
US6031091A (en) * 1987-09-21 2000-02-29 Gen-Probe Incorporated Non-nucleotide linking reagents for nucleotide probes
US5585481A (en) * 1987-09-21 1996-12-17 Gen-Probe Incorporated Linking reagents for nucleotide probes
US5998135A (en) * 1989-02-24 1999-12-07 Enzo Diagnostics, Inc. Energy transfer hybridization assay using intercalators and lanthanide metals
AU7579991A (en) * 1990-02-20 1991-09-18 Gilead Sciences, Inc. Pseudonucleosides and pseudonucleotides and their polymers
US5332659A (en) * 1992-04-09 1994-07-26 The United States Of America As Represented By The Secretary Of The Navy Light emission-or absorbance-based binding assays for polynucleic acids
US5886177A (en) * 1994-01-11 1999-03-23 Isis Pharmaceuticals, Inc. Phosphate linked oligomers
GB9604669D0 (en) * 1996-03-05 1996-05-01 Ciba Geigy Ag Chemical compounds
CA2252048C (en) * 1996-04-12 2008-03-11 The Public Health Research Institute Of The City Of New York, Inc. Detection probes, kits and assays
ATE275573T1 (en) * 1996-05-15 2004-09-15 Univ Rochester NEW NUCLEOSIDE ANALOGUES THAT HAVE POLYCYCLIC AROMATIC GROUPS AS SUBSTITUENTS, METHOD FOR THEIR PRODUCTION AND THEIR APPLICATIONS
US6248518B1 (en) * 1996-10-29 2001-06-19 Board Of Regents Of University Of Nebraska Method for detecting point mutations in DNA utilizing fluorescence energy transfer
US6172209B1 (en) * 1997-02-14 2001-01-09 Isis Pharmaceuticals Inc. Aminooxy-modified oligonucleotides and methods for making same
US6117973A (en) * 1997-02-24 2000-09-12 Georgia Tech Research Corp. PNA monomers with electron donor or acceptor
US6506567B2 (en) * 2000-01-31 2003-01-14 Fuji Photo Film Co., Ltd. Water-soluble flourescent intercalator compound
US20030165837A1 (en) * 2000-06-14 2003-09-04 The Scripps Research Institute Method for screening DNA binding

Also Published As

Publication number Publication date
CN1653079A (en) 2005-08-10
WO2003052133A2 (en) 2003-06-26
AU2002358464B2 (en) 2006-04-27
AU2002358463A1 (en) 2003-06-30
WO2003051901A3 (en) 2003-11-27
AU2002358463A8 (en) 2003-06-30
WO2003052132A2 (en) 2003-06-26
AU2002358462A1 (en) 2003-06-30
CN1653079B (en) 2010-06-16
WO2003052134A2 (en) 2003-06-26
AU2002361946A1 (en) 2003-06-30
AU2002358464A1 (en) 2003-06-30
AU2002358462A8 (en) 2003-06-30
WO2003052134A3 (en) 2004-03-25
WO2003052133A3 (en) 2003-10-02
WO2003051901A2 (en) 2003-06-26
WO2003052132A3 (en) 2003-10-09
EP1468007A2 (en) 2004-10-20

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Legal Events

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MK6 Application lapsed section 142(2)(f)/reg. 8.3(3) - pct applic. not entering national phase