JP2006525022A5 - - Google Patents
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- JP2006525022A5 JP2006525022A5 JP2006513381A JP2006513381A JP2006525022A5 JP 2006525022 A5 JP2006525022 A5 JP 2006525022A5 JP 2006513381 A JP2006513381 A JP 2006513381A JP 2006513381 A JP2006513381 A JP 2006513381A JP 2006525022 A5 JP2006525022 A5 JP 2006525022A5
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Families Citing this family (49)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7875440B2 (en) | 1998-05-01 | 2011-01-25 | Arizona Board Of Regents | Method of determining the nucleotide sequence of oligonucleotides and DNA molecules |
| US6780591B2 (en) * | 1998-05-01 | 2004-08-24 | Arizona Board Of Regents | Method of determining the nucleotide sequence of oligonucleotides and DNA molecules |
| US7501245B2 (en) * | 1999-06-28 | 2009-03-10 | Helicos Biosciences Corp. | Methods and apparatuses for analyzing polynucleotide sequences |
| US6818395B1 (en) * | 1999-06-28 | 2004-11-16 | California Institute Of Technology | Methods and apparatus for analyzing polynucleotide sequences |
| EP1368497A4 (en) | 2001-03-12 | 2007-08-15 | California Inst Of Techn | METHODS AND APPARATUS FOR ANALYZING ASYNCHRONOUS BASE EXTENSION POLYNUCLEOTIDE SEQUENCES |
| US9261460B2 (en) * | 2002-03-12 | 2016-02-16 | Enzo Life Sciences, Inc. | Real-time nucleic acid detection processes and compositions |
| DE10154317B4 (de) * | 2001-10-26 | 2005-06-09 | Epigenomics Ag | Verfahren zum Nachweis von Cytosin-Methylierungen in immobilisierten DNA Proben |
| US20040054162A1 (en) * | 2001-10-30 | 2004-03-18 | Hanna Michelle M. | Molecular detection systems utilizing reiterative oligonucleotide synthesis |
| US7045319B2 (en) * | 2001-10-30 | 2006-05-16 | Ribomed Biotechnologies, Inc. | Molecular detection systems utilizing reiterative oligonucleotide synthesis |
| US9353405B2 (en) | 2002-03-12 | 2016-05-31 | Enzo Life Sciences, Inc. | Optimized real time nucleic acid detection processes |
| US20050170367A1 (en) * | 2003-06-10 | 2005-08-04 | Quake Stephen R. | Fluorescently labeled nucleoside triphosphates and analogs thereof for sequencing nucleic acids |
| WO2005042709A2 (en) * | 2003-10-29 | 2005-05-12 | Ribomed Biotechnologies, Inc. | Compositions, methods and detection technologies for reiterative oligonucleotide synthesis |
| US7169560B2 (en) | 2003-11-12 | 2007-01-30 | Helicos Biosciences Corporation | Short cycle methods for sequencing polynucleotides |
| US20060172408A1 (en) * | 2003-12-01 | 2006-08-03 | Quake Steven R | Device for immobilizing chemical and biochemical species and methods of using same |
| WO2005080605A2 (en) | 2004-02-19 | 2005-09-01 | Helicos Biosciences Corporation | Methods and kits for analyzing polynucleotide sequences |
| US20050239085A1 (en) * | 2004-04-23 | 2005-10-27 | Buzby Philip R | Methods for nucleic acid sequence determination |
| US20050260609A1 (en) * | 2004-05-24 | 2005-11-24 | Lapidus Stanley N | Methods and devices for sequencing nucleic acids |
| US20070117104A1 (en) * | 2005-11-22 | 2007-05-24 | Buzby Philip R | Nucleotide analogs |
| US7476734B2 (en) * | 2005-12-06 | 2009-01-13 | Helicos Biosciences Corporation | Nucleotide analogs |
| DE602005027700D1 (de) | 2004-05-25 | 2011-06-09 | Helicos Biosciences Corp | Verfahren zur nukleinsäureimmobilisierung |
| US20070117103A1 (en) * | 2005-11-22 | 2007-05-24 | Buzby Philip R | Nucleotide analogs |
| US20060024678A1 (en) * | 2004-07-28 | 2006-02-02 | Helicos Biosciences Corporation | Use of single-stranded nucleic acid binding proteins in sequencing |
| WO2006037102A2 (en) * | 2004-09-27 | 2006-04-06 | Epicentre Technologies | Methods for identifying polymerase inhibitors |
| WO2006036943A2 (en) | 2004-09-27 | 2006-04-06 | The University Of North Carolina At Chapel Hill | Determination of molecular age by detection of ink4a/arf expression |
| US20060118754A1 (en) * | 2004-12-08 | 2006-06-08 | Lapen Daniel C | Stabilizing a polyelectrolyte multilayer |
| US7220549B2 (en) | 2004-12-30 | 2007-05-22 | Helicos Biosciences Corporation | Stabilizing a nucleic acid for nucleic acid sequencing |
| US20060172328A1 (en) * | 2005-01-05 | 2006-08-03 | Buzby Philip R | Methods and compositions for correcting misincorporation in a nucleic acid synthesis reaction |
| US7482120B2 (en) * | 2005-01-28 | 2009-01-27 | Helicos Biosciences Corporation | Methods and compositions for improving fidelity in a nucleic acid synthesis reaction |
| US20060263790A1 (en) * | 2005-05-20 | 2006-11-23 | Timothy Harris | Methods for improving fidelity in a nucleic acid synthesis reaction |
| DE602006018622D1 (de) * | 2005-06-30 | 2011-01-13 | Ge Healthcare Bio Sciences | Nachweisverfahren für die genexpression |
| US20070031875A1 (en) * | 2005-08-05 | 2007-02-08 | Helicos Biosciences Corporation | Signal pattern compositions and methods |
| US7666593B2 (en) | 2005-08-26 | 2010-02-23 | Helicos Biosciences Corporation | Single molecule sequencing of captured nucleic acids |
| US20070117102A1 (en) * | 2005-11-22 | 2007-05-24 | Buzby Philip R | Nucleotide analogs |
| US20070128610A1 (en) * | 2005-12-02 | 2007-06-07 | Buzby Philip R | Sample preparation method and apparatus for nucleic acid sequencing |
| US20080309926A1 (en) * | 2006-03-08 | 2008-12-18 | Aaron Weber | Systems and methods for reducing detected intensity non uniformity in a laser beam |
| US7397546B2 (en) * | 2006-03-08 | 2008-07-08 | Helicos Biosciences Corporation | Systems and methods for reducing detected intensity non-uniformity in a laser beam |
| CA2724343A1 (en) | 2008-05-15 | 2009-11-19 | Ribomed Biotechnologies, Inc. | Methods and reagents for detecting cpg methylation with a methyl cpg binding protein (mbp) |
| WO2010009060A2 (en) * | 2008-07-13 | 2010-01-21 | Ribomed Biotechnologies, Inc. | Molecular beacon-based methods for detection of targets using abscription |
| RU2011141720A (ru) | 2009-03-15 | 2013-04-27 | Рибомед Байотекнолоджиз, Инк. | Молекулярное обнаружение, основанное на абскрипции |
| WO2011034895A1 (en) * | 2009-09-15 | 2011-03-24 | Regents Of The University Of Colorado | Compositions, methods and uses for nucleotide analogues |
| US8809531B2 (en) * | 2010-04-02 | 2014-08-19 | Pharmacophotonics, Inc. | Rhodamine dyes and conjugates |
| ES3018861T3 (en) * | 2011-12-13 | 2025-05-19 | Univ Oslo Hf | Method for detection of hydroxymethylation status |
| WO2015164602A2 (en) * | 2014-04-23 | 2015-10-29 | Children's Medical Center Corporation | High-throughput structure determination using nucleic acid calipers |
| CA3069985A1 (en) * | 2017-07-24 | 2019-01-31 | Quantum-Si Incorporated | High intensity labeled reactant compositions and methods for sequencing |
| CN108467889A (zh) * | 2018-04-04 | 2018-08-31 | 苏州创澜生物科技有限公司 | 一种用于多重pcr的荧光探针及其应用 |
| AU2019300169A1 (en) | 2018-07-13 | 2021-01-28 | Quantum-Si Incorporated | Biconjugatable labels and methods of use |
| MX2021008867A (es) | 2019-01-23 | 2021-08-19 | Quantum Si Inc | Composiciones reactantes marcadas de intensidad alta y metodos de secuenciamiento. |
| US11591629B2 (en) * | 2019-12-23 | 2023-02-28 | Cheng-Yao Chen | Method and kit for template-independent nucleic acid synthesis |
| WO2022023753A1 (en) | 2020-07-30 | 2022-02-03 | Cambridge Epigenetix Limited | Compositions and methods for nucleic acid analysis |
Family Cites Families (42)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3996345A (en) * | 1974-08-12 | 1976-12-07 | Syva Company | Fluorescence quenching with immunological pairs in immunoassays |
| US4351760A (en) * | 1979-09-07 | 1982-09-28 | Syva Company | Novel alkyl substituted fluorescent compounds and polyamino acid conjugates |
| US4582788A (en) * | 1982-01-22 | 1986-04-15 | Cetus Corporation | HLA typing method and cDNA probes used therein |
| US4786600A (en) * | 1984-05-25 | 1988-11-22 | The Trustees Of Columbia University In The City Of New York | Autocatalytic replication of recombinant RNA |
| US5503979A (en) * | 1984-05-25 | 1996-04-02 | The Trustees Of Columbia University In The City Of New York | Method of using replicatable hybridzable recombinant RNA probes |
| US4683194A (en) * | 1984-05-29 | 1987-07-28 | Cetus Corporation | Method for detection of polymorphic restriction sites and nucleic acid sequences |
| US4739044A (en) * | 1985-06-13 | 1988-04-19 | Amgen | Method for derivitization of polynucleotides |
| US4757141A (en) * | 1985-08-26 | 1988-07-12 | Applied Biosystems, Incorporated | Amino-derivatized phosphite and phosphate linking agents, phosphoramidite precursors, and useful conjugates thereof |
| US5130238A (en) * | 1988-06-24 | 1992-07-14 | Cangene Corporation | Enhanced nucleic acid amplification process |
| US5508178A (en) * | 1989-01-19 | 1996-04-16 | Rose; Samuel | Nucleic acid amplification using single primer |
| US5766849A (en) * | 1989-07-11 | 1998-06-16 | Gen-Probe Incorporated | Methods of amplifying nucleic acids using promoter-containing primer sequence |
| CA2020958C (en) * | 1989-07-11 | 2005-01-11 | Daniel L. Kacian | Nucleic acid sequence amplification methods |
| US6114513A (en) * | 1990-01-11 | 2000-09-05 | Isis Pharmaceuticals, Inc. | Thiol-derivatized oligonucleotides |
| US5578718A (en) * | 1990-01-11 | 1996-11-26 | Isis Pharmaceuticals, Inc. | Thiol-derivatized nucleosides |
| US5194370A (en) * | 1990-05-16 | 1993-03-16 | Life Technologies, Inc. | Promoter ligation activated transcription amplification of nucleic acid sequences |
| US5595891A (en) * | 1990-07-19 | 1997-01-21 | Behringwerke Ag | Method for producing a polynucleotide for use in single primer amplification |
| US5888819A (en) * | 1991-03-05 | 1999-03-30 | Molecular Tool, Inc. | Method for determining nucleotide identity through primer extension |
| US5169766A (en) * | 1991-06-14 | 1992-12-08 | Life Technologies, Inc. | Amplification of nucleic acid molecules |
| JP4137996B2 (ja) * | 1992-05-06 | 2008-08-20 | ジェン−プローブ・インコーポレイテッド | 核酸配列増幅方法,組成物及びキット |
| WO1994001445A1 (en) * | 1992-07-14 | 1994-01-20 | Research Corporation Technologies, Inc. | Base-protected nucleotide analogs with protected thiol groups |
| DE69310179T2 (de) * | 1992-07-31 | 1997-07-31 | Behringwerke Ag | Verfahren zur einführung von definierten sequenzen am 3' ende von polynukleotiden |
| ZA936016B (en) * | 1992-08-24 | 1994-03-10 | Akzo Nv | Method for nucleic acid amplification |
| US5571669A (en) * | 1993-01-14 | 1996-11-05 | The University Of Pennsylvania | Transcription and nucleic acid sequence determination with short primer DNA/RNA molecules and RNA polymerase |
| US5597694A (en) * | 1993-10-07 | 1997-01-28 | Massachusetts Institute Of Technology | Interspersed repetitive element-bubble amplification of nucleic acids |
| US5648211A (en) * | 1994-04-18 | 1997-07-15 | Becton, Dickinson And Company | Strand displacement amplification using thermophilic enzymes |
| FR2724934B1 (fr) * | 1994-09-26 | 1997-01-24 | Bio Merieux | Oligonucleotide chimere et son utilisation dans l'obtention de transcrits d'un acide nucleique |
| US5684142A (en) * | 1995-06-07 | 1997-11-04 | Oncor, Inc. | Modified nucleotides for nucleic acid labeling |
| FR2737223B1 (fr) * | 1995-07-24 | 1997-09-12 | Bio Merieux | Procede d'amplification de sequences d'acide nucleique par deplacement, a l'aide d'amorces chimeres |
| US5912340A (en) * | 1995-10-04 | 1999-06-15 | Epoch Pharmaceuticals, Inc. | Selective binding complementary oligonucleotides |
| AU3891097A (en) * | 1996-07-24 | 1998-02-10 | Michelle M. Hanna | Base-protected nucleotide analogs with protected thiol groups |
| US5849546A (en) * | 1996-09-13 | 1998-12-15 | Epicentre Technologies Corporation | Methods for using mutant RNA polymerases with reduced discrimination between non-canonical and canonical nucleoside triphosphates |
| US5846723A (en) * | 1996-12-20 | 1998-12-08 | Geron Corporation | Methods for detecting the RNA component of telomerase |
| US5858801A (en) * | 1997-03-13 | 1999-01-12 | The United States Of America As Represented By The Secretary Of The Navy | Patterning antibodies on a surface |
| US6251594B1 (en) * | 1997-06-09 | 2001-06-26 | Usc/Norris Comprehensive Cancer Ctr. | Cancer diagnostic method based upon DNA methylation differences |
| US6114519A (en) * | 1997-10-15 | 2000-09-05 | Isis Pharmaceuticals, Inc. | Synthesis of sulfurized oligonucleotides |
| US6268131B1 (en) * | 1997-12-15 | 2001-07-31 | Sequenom, Inc. | Mass spectrometric methods for sequencing nucleic acids |
| US6803485B2 (en) * | 1999-02-26 | 2004-10-12 | Bracco Imaging S.P.A. | Process for the preparation of iopamidol |
| WO2001020017A2 (en) * | 1999-09-14 | 2001-03-22 | Yeda Research And Development Co. Ltd. | Metal cluster containing nucleotides and nucleic acids, and intermediates therefor |
| US20020168641A1 (en) * | 2001-03-09 | 2002-11-14 | Bruce Mortensen | Fluorescein-cyanine 5 as a fluorescence resonance energy transfer pair |
| US20040054162A1 (en) * | 2001-10-30 | 2004-03-18 | Hanna Michelle M. | Molecular detection systems utilizing reiterative oligonucleotide synthesis |
| US7045319B2 (en) * | 2001-10-30 | 2006-05-16 | Ribomed Biotechnologies, Inc. | Molecular detection systems utilizing reiterative oligonucleotide synthesis |
| WO2005042709A2 (en) * | 2003-10-29 | 2005-05-12 | Ribomed Biotechnologies, Inc. | Compositions, methods and detection technologies for reiterative oligonucleotide synthesis |
-
2003
- 2003-04-29 US US10/425,037 patent/US20040054162A1/en not_active Abandoned
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2004
- 2004-04-29 AU AU2004235368A patent/AU2004235368A1/en not_active Abandoned
- 2004-04-29 CA CA002523442A patent/CA2523442A1/en not_active Abandoned
- 2004-04-29 JP JP2006513381A patent/JP2006525022A/ja active Pending
- 2004-04-29 US US10/551,775 patent/US20060204964A1/en not_active Abandoned
- 2004-04-29 WO PCT/US2004/013031 patent/WO2004096997A2/en not_active Ceased
- 2004-04-29 EP EP04750780A patent/EP1622923A4/en not_active Withdrawn
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