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31. 发明公开

EP1179085A4 METHOD FOR SEQUENCING NUCLEIC ACID MOLECULES 有权
标题翻译：方法用于测序核酸分子的
公开(公告)号：EP1179085A4
公开(公告)日：2004-11-10
申请号：EP00932575
申请日：2000-05-18
申请人： CORNELL RES FOUNDATION INC
发明人： KORLACH JONAS , WEBB WATT W , LEVENE MICHAEL , TURNER STEPHEN , CRAIGHEAD HAROLD G , FOQUET MATHIEU
IPC分类号： G01N33/50 , C12M1/00 , C12N15/09 , C12Q1/68 , G01N33/58 , C12P19/34 , C12M1/34
CPC分类号： C12Q1/6874 , C12Q1/6869 , Y10S436/80 , Y10S436/805 , Y10T436/143333 , C12Q2565/537 , C12Q2537/149 , C12Q2561/113 , C12Q2521/543 , C12Q2565/518 , C12Q2561/12
摘要： The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the templage nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.

32. 发明公开

EP1389241A2 POLYNUCLEOTIDE SEQUENCING METHOD 有权
标题翻译：用于多核苷酸测序的方法
公开(公告)号：EP1389241A2
公开(公告)日：2004-02-18
申请号：EP02727736.7
申请日：2002-05-20
申请人： Medical Biosystems Ltd.
发明人： DENSHAM, Daniel Henry
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q2565/632 , C12Q2533/101 , C12Q2521/543
摘要： A method for determining the sequence of a polynucleotide comprising the steps of (i) contacting a polynucleotide processive enzyme immobilised in a fixed position, with a target polynucleotide under conditions sufficient to induce enzyme activity; (ii) detecting an effect consequent on the interaction of the enzyme and the polynucleotide, wherein the effect is detected by measurement of a non-linear optical signal or a linear signal coupled to a non-linear signal.

33. 发明公开

EP1180169A1 METHODS OF DETECTING POLYNUCLEOTIDE KINASE AND ITS USE AS A LABEL 有权
标题翻译：检测方法的多核苷酸及其用途为指标
公开(公告)号：EP1180169A1
公开(公告)日：2002-02-20
申请号：EP01920491.6
申请日：2001-03-23
申请人： LUMIGEN, INC.
发明人： AKHAVAN-TAFTI, Hashem
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/00
CPC分类号： C12Q1/485 , C12Q1/682 , C12Q2533/107 , C12Q2521/543 , C12Q2521/501
摘要： Methods of detecting or measuring the activity of polynucleotide kinase are disclosed as well as methods of detecting an analyte in an assay using polynucleotide kinase as a label on a member of a specific binding pair. The methods rely on the phosphorylation of an oligonucleotide followed by ligation of the oligonucleotide 5'-phosphate onto another template-bound oligonucleotide. The presence of the ligated product signals the presence of polynucleotide kinase. In preferred embodiments, phosphorylation of an oligonucleotide enables the consecutive ligation of a set of oligonucleotides. The oligonucleotides so ligated can be detectably labeled with, for example, other enzymes to provide highly sensitive detection methods.

34. 发明公开

EP0689610A1 DNA SEQUENCING BY MASS SPECTROMETRY VIA EXONUCLEASE DEGRADATION 失效
标题翻译：质量对核酸外切酶的脱除方法的DNA序列识别
公开(公告)号：EP0689610A1
公开(公告)日：1996-01-03
申请号：EP94911643.0
申请日：1994-03-18
申请人： SEQUENOM, INC.
发明人： Köster, Hubert
IPC分类号： G01N33 , C07F9 , C07H19 , C07H21 , C12N15 , C12Q1 , G01N35
CPC分类号： C12Q1/6872 , C07F9/2408 , C07F9/2429 , C07H19/06 , C07H19/10 , C07H19/16 , C07H19/20 , C07H21/00 , G01N35/1067 , C12Q2565/631 , C12Q2537/157 , C12Q2521/543 , C12Q2521/325 , C12Q2537/143 , C12Q2525/117 , C12Q2521/319
摘要： Methods for determining the sequence of nucleic acids by cleaving the nucleic acid unilaterally from a first end with an exonuclease activity to sequentially release individual nucleotides, identifying each of the sequentially release nucleotides by mass spectometry, and determining the sequence of the nucleic acid from the identified nucleotides are disclosed. The method is amenable to multiplexing for simutaneously determining more than one nucleic acid sequence.

35. 发明公开

EP0243797A2 Method for amplifying enzyme activity, enzyme conjugates suitable therefor and their preparation 无效
标题翻译：一种用于提高酶的活性的方法，化酶共轭体适合于该目的和它们的制备。
公开(公告)号：EP0243797A2
公开(公告)日：1987-11-04
申请号：EP87105569.5
申请日：1987-04-15
申请人： NORTHEASTERN UNIVERSITY
发明人： Giese, Roger W. , Ehrat, Markus , Cecchini, Douglas J.
IPC分类号： C12Q1/00 , C12Q1/68
CPC分类号： C12Q1/68 , C12Q1/00 , Y10S435/962 , Y10S435/966 , C12Q2521/543 , C12Q2565/518
摘要： The invention relates to a method for amplifying enzyme activity, spcecific enzyme conjugates suitable therefor and a method for their preparation.
Enzyme amplification is achieved by covalently bonding enzyme to a support material via a molecular chain which is a substrate for the enzyme, then introducing a small amount of free enzyme to this system, causing release of a large amount of bound enzyme. Alternatively, complementary enzymatically inactive fragments of an active enzyme, which can recombine to form active enzyme, are covalently attached to separate support materials by a molecular chain material which is a substrate for the active enzyme, and these two fragment-support conjugates are connected in series. In a second alternative embodiment, two different active enzymes may be used. The method is suited e.g. for the S-peptide and S-protein fragments of ribonuclease S (RNaseS) which are preferably coupled to Sepharose gels, via a polycytidylic acid substrate leash. Also the released fragments recombine to give RNaseS activity. Thus this system provides more enzymatic activity from the system than is applied. 20 hours to yield activity equivalent to 1.911-g of RNase, by applying it to the S-peptide gel followed by combination of the released S-peptide with excess S-protein. Also, application of 1 ng of RNaseA to a three-stage amplification system in which each stage consists of a pair of S-peptide and S-protein gels produced cumulative amplifications of 4, 9, 52 and 25x after each to the stages, respectively. Only 1 - 2 mg amounts of each gel are used per stage in these experiments, reflecting the significant production of RNaseS activity.
Molecular amplification factors of -2.104 can be achieved.
摘要翻译：本发明涉及的方法用于扩增酶活性，spcecific酶偶联物适合于此的方法和用于它们的制备。酶扩增通过共价到载体材料经由一个分子链中的所有其是酶的底物结合的酶，然后引入游离酶少量该系统，引起大量的结合的酶的释放来实现。可替代地，活性酶，其可重新结合形成活性酶的互补无酶活性的片段共价由分子链材料的所有其对于活性酶的底物附着到单独的载体材料，和论文2片段 - 载体缀合物被连接在系列。在第二替代实施例中，可以使用两种不同的活性酶。该方法是适合E.G. 用于S-肽和核糖核酸酶S-（RNA酶），其优选地耦合到琼脂糖凝胶的S-蛋白质片段，通过胞苷酸基板皮带。因此，释放的片段重组给予核糖核酸酶和活性。因此，该系统提供了从系统比应用更酶活性。 20小时，以得到同等活性1.9亩克RNA酶的，通过将其应用到随后用过量S-蛋白释放的S-肽的组合的S-肽凝胶。所以，分别应用1纳克的RNase A的，其中一对S-肽和S-蛋白凝胶中的每一级besteht产生的4，图9，52和25倍累积扩增每个所述阶段之后，一个三阶段的放大系统，每个凝胶的只有1〜2毫克量以每级中使用在合成实验，反映了显著生产RNA酶和活性。的相似2.10分子扩增因子<4>可以实现的。

36. 发明授权

EP2158476B1 CHEMICAL FUNCTIONALIZATION OF SOLID-STATE NANOPORES AND NANOPORE ARRAYS AND APPLICATIONS THEREOF 有权转让
公开(公告)号：EP2158476B1
公开(公告)日：2019-04-24
申请号：EP08827096.2
申请日：2008-05-08
申请人： The Trustees of the Boston University
发明人： MELLER, Amit , WANUNU, Meni
IPC分类号： G01N27/00 , C12Q1/68 , B81C1/00
CPC分类号： G01N33/48721 , B82Y15/00 , C12Q1/6869 , G01N21/6428 , G01N2021/6439 , Y10T428/2982 , Y10T436/143333 , C12Q2521/543 , C12Q2565/631
摘要： Chemical functionalization of solid-state nanopores and nanopore arrays and applications thereof. Nanopores are extremely sensitive single-molecule sensors. Recently, electron beams have been used to fabricate synthetic nanopores in thin solid-state membranes with sub-nanometer resolution. A new class of chemically modified nanopore sensors are provided with two approaches for monolayer coating of nanopores by: (1) self-assembly from solution, in which nanopores −10 nm diameter can be reproductibly coated, and (2) self-assembly under voltage-driven electrolyte flow, in which 5 nm nanopores may be coated. Applications of chemically modified nanopore are provided including: the detection of biopolymers such as DNA and RNA; immobilizing enzymes or other proteins for detection or for generating chemical gradients; and localized pH sensing.

37. 发明公开

EP3365273A1 USE OF FLUOROPOLYMERS AS A HYDROPHOBIC LAYER TO SUPPORT LIPID BILAYER FORMATION FOR NANOPORE 审中-公开
公开(公告)号：EP3365273A1
公开(公告)日：2018-08-29
申请号：EP16858352.4
申请日：2016-10-21
申请人： F. Hoffmann-La Roche AG , Roche Diagnostics GmbH
发明人： FOSTER, John , HENCK, Steven
IPC分类号： B82Y15/00 , B82Y40/00 , C12N15/09 , C12Q1/68 , G01N27/26 , G01N27/327 , G01N33/58
CPC分类号： G01N27/3278 , B82Y15/00 , C12Q1/6869 , G01N27/3276 , G01N27/40 , G01N33/48721 , C12Q2535/122 , C12Q2563/116 , C12Q2565/607 , C12Q2565/631 , C12Q2521/543
摘要： A method of sequencing a DNA sample is disclosed. A nanopore-based sequencing device is provided. The nanopore-based sequencing device includes a conductive layer. The device further includes a working electrode disposed above the conductive layer. The device further includes a side wall disposed above the working electrode, wherein the side wall and the working electrode form a well in which an electrolyte may be contained, and wherein at least an upper portion of the side wall comprises a hydrophobic portion formed by a fluoropolymer material. The DNA sample is sequenced using the nanopore-based sequencing device.

38. 发明公开

EP3294908A1 FIELD-EFFECT APPARATUS AND METHODS FOR SEQUENCING NUCLEIC ACIDS 有权
公开(公告)号：EP3294908A1
公开(公告)日：2018-03-21
申请号：EP16727012
申请日：2016-05-11
申请人： ILLUMINA INC , UNIV CALIFORNIA
发明人： GUNDERSON KEVIN L , BAI JINGWEI , CHEN CHENG YAO , MANDELL JEFFREY G , PEISAJOVICH SERGIO , BOYANOV BOYAN , COLLINS PHILIP G , WEISS GREGORY A
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12Q2521/543 , C12Q2565/607
摘要： The present disclosure provides a method for sequencing nucleic acids. The method can include polymerase catalyzed incorporation of nucleotides into a nascent nucleic acid strand against a nucleic acid template, wherein the polymerase is attached to a charge sensor that detects nucleotide incorporation events. One or more non-natural nucleotide types that each produce a unique signatures at the charge sensor can be used to uniquely identify different nucleotides in the template nucleic acid.

39. 发明公开

EP3107865A4 FLUORESCENCE-BASED ANALYSIS OF BIOPOLYMERS USING NANOPORES 审中-公开
公开(公告)号：EP3107865A4
公开(公告)日：2017-10-18
申请号：EP15751896
申请日：2015-02-23
申请人： UNIV NORTHEASTERN
发明人： IVANKIN ANDREY , LARKIN JOSEPH , HENLEY ROBERT , WANUNU MENI
IPC分类号： G01N21/64 , C12Q1/68 , G01N21/77 , G01N33/487
CPC分类号： C12Q1/6869 , G01N21/6408 , G01N21/6428 , G01N21/6458 , G01N33/48721 , G01N2021/6439 , G01N2021/7786 , C12Q2521/543 , C12Q2563/137 , C12Q2565/631

40. 发明授权

EP2675917B1 METHODS AND COMPOSITIONS FOR THE TARGET-LOCALIZED ANCHORING OF DETECTABLE LABEL 有权
标题翻译：用于可检测标签的目标本地化锚定的方法和组合物
公开(公告)号：EP2675917B1
公开(公告)日：2017-09-13
申请号：EP12746537.5
申请日：2012-02-14
申请人： Headway Technologies, Inc.
发明人： HU, Celine , PERKINS, Julie , GAO, Hetian , WANG, Lisen
IPC分类号： C12Q1/68 , C07D495/04 , C07D207/408
CPC分类号： C12Q1/6816 , C07D207/408 , C07D495/04 , C12Q1/6834 , C12Q2537/125 , C12Q2565/519 , C12Q2521/543 , C12Q2525/197 , C12Q2563/131 , C12Q2563/143 , C12Q2563/149

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