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31. 发明申请

WO2003094101A1 RAY-BASED IMAGE ANALYSIS FOR BIOLOGICAL SPECIMENS 审中-公开
标题翻译：基于RAY的生物样本图像分析
公开(公告)号：WO2003094101A1
公开(公告)日：2003-11-13
申请号：PCT/US2003/013500
申请日：2003-04-29
申请人： AMERSHAM BIOSCIENCES CORP.
发明人： HANSEN, Richard, L. , KARSH, William, J.
IPC分类号： G06K9/00
CPC分类号： G06K9/50 , G06K9/0014 , G06T7/0012 , G06T2207/30024
摘要： Image acquisition and analysis systems and methods are provided. A ray-based approach may be used to process images for cell-based assays. Such cell-based assays may be used to evaluate drugs or other compounds or to perform other biological studies. A scanning laser microscope or other equipment may be used to gather image data from fluorescently-marked cells (42) or other suitable specimens. The rays (40) are radially-oriented with respect to the cell nuclei (44). Seed points (48) within the nuclei (44) may be identified. The rays (40) may extend outward from the seed points (48) or other suitable ray origins until the rays (40) are terminated according to ray termination criteria. The intensity of the image data that is associated with each of the rays (40) may be analyzed to generate various parameters. For example, a peak intensity of the image data along each ray may be identified. Statistical calculations may be performed.
摘要翻译：提供了图像采集和分析系统和方法。基于射线的方法可用于处理基于细胞的测定的图像。这种基于细胞的测定可用于评估药物或其他化合物或进行其他生物学研究。扫描激光显微镜或其他设备可用于从荧光标记的细胞（42）或其他合适的样品中收集图像数据。射线（40）相对于细胞核（44）径向取向。可以鉴定细胞核（44）内的种子点（48）。射线（40）可以从种子点（48）或其他合适的射线起点向外延伸，直到根据射线终止标准终止射线（40）。可以分析与每个射线（40）相关联的图像数据的强度，以生成各种参数。例如，可以识别沿着每个射线的图像数据的峰值强度。可以进行统计学计算。

32. 发明申请

WO0125485A3 METHOD FOR DETECTING MUTATIONS USING ARRAYED PRIMER EXTENSION 审中-公开
标题翻译：使用阵列扩展来检测突变的方法
公开(公告)号：WO0125485A3
公开(公告)日：2002-09-12
申请号：PCT/US0027319
申请日：2000-10-04
申请人： AMERSHAM BIOSCIENCES CORP , ZHAO XIAODONG
发明人： ZHAO XIAODONG
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q1/6874 , C12Q2565/537 , C12Q2565/525 , C12Q2535/125 , C12Q2535/101
摘要： A method for detecting a mutation in a target nucleic acid sequence that comprises attaching oligonucleotide primers to a substrate, wherein the oligonucleotide primers have a sequence that is complementary to the target nucleic acid sequence, and wherein the oligonucleotide primers are grouped according to the identity of the first base which would be expected to be added to the primer through the process of primer extension; hybridizing to the oligonucleotide primers a sample nucleic acid sequence which possibly contains a mutation; extending each oligonucleotide primer by one base using a reaction mixture comprising labeled ddNTPs; and detecting a mutation in the sample nucleic acid sequence by detecting the presence of a labeled ddNTP which does not correspond to the identity of the base expected to be added to the primer through the process of primer extension. Mutations may be detected in mRNA or DNA. The labeled ddNTPs may be labeled with a fluorescent dye, a chemiluminescent reagent, a radioactive label, or an electrically conductive tag. A nucleic acid array and a kit for detecting genetic mutations are also disclosed. The instant disclosure also pertains to a method for detecting a mutation in a target nucleic acid sequence that comprises identifying the base expected to be added to a primer located at a particular coordinate on an oligonucleotide array as expected from the target nucleic acid sequence; identifying the base actually added to the primer located at the particular coordinate on the oligonucleotide array through the process of primer extension; comparing the base actually added to the primer at the particular coordinate with the base expected to be added to the primer at the particular coordinate; and reporting those instances where the bases are not the same, in order to identify a mutation.
摘要翻译：一种检测靶核酸序列突变的方法，包括将寡核苷酸引物连接到底物上，其中所述寡核苷酸引物具有与靶核酸序列互补的序列，并且其中所述寡核苷酸引物根据通过引物延伸的过程将预期添加到引物中的第一个碱基; 与寡核苷酸引物杂交可能含有突变的样品核酸序列; 使用包含标记的ddNTP的反应混合物将每个寡核苷酸引物延伸一个碱基; 并通过检测通过引物延伸的过程检测不符合预期添加到引物的碱基的同一性的标记的ddNTP的存在来检测样品核酸序列中的突变。可以在mRNA或DNA中检测突变。标记的ddNTP可以用荧光染料，化学发光试剂，放射性标记或导电标签进行标记。还公开了核酸阵列和用于检测遗传突变的试剂盒。本公开还涉及用于检测靶核酸序列中的突变的方法，其包括鉴定从靶核酸序列预期的预期添加到位于寡核苷酸阵列上的特定坐标的引物的碱基; 通过引物延伸鉴定实际添加到位于寡核苷酸阵列上特定座标上的引物的碱基; 将在特定坐标处实际添加到底漆的碱与预期在特定坐标处加入底漆的碱进行比较; 并报告那些基因不相同的情况，以便鉴定突变。

33. 发明申请

WO0177658A3 CASSETTE FOR BUFFERLESS GEL ELECTROPHORESIS 审中-公开
标题翻译：无溶胶凝胶电泳
公开(公告)号：WO0177658A3
公开(公告)日：2002-09-06
申请号：PCT/US0111600
申请日：2001-04-09
申请人： AMERSHAM BIOSCIENCES CORP , READ DOUG , HENNESSY LORI , ISLAM MOHAMMED REZAUL
发明人： READ DOUG , HENNESSY LORI , ISLAM MOHAMMED REZAUL
IPC分类号： G01N27/447
CPC分类号： G01N27/44704 , G01N27/44743
摘要： A cassette and electrophoretic gel assembly includes a non-conductive cassette, two solid buffer reservoirs, and an agarose gel. The assembly is disposable, and the sample wells on the gel are in standard microtiter plate format. The configuration is such that the gel is continuously in electrical contact with the electrodes in operation despite the production/migration of water and other exudates.
摘要翻译：盒和电泳凝胶组件包括非导电盒，两个固体缓冲液储存器和琼脂糖凝胶。组件是一次性的，凝胶上的样品孔是标准微量滴定板形式。该配置使得尽管水和其它渗出物的生成/迁移，凝胶在操作中仍与电极连续地电接触。

34. 发明申请

WO2005123957A2 METHOD FOR NUCLEIC ACID ANALYSIS 审中-公开
标题翻译：核酸分析方法
公开(公告)号：WO2005123957A2
公开(公告)日：2005-12-29
申请号：PCT/US2005/020378
申请日：2005-06-09
申请人： AMERSHAM BIOSCIENCES CORP , FULLER, Carl, W. , NELSON, John, R.
发明人： FULLER, Carl, W. , NELSON, John, R.
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6834 , C12Q1/6869 , C12Q2565/519 , C12Q2565/301 , C12Q2563/137 , C12Q2527/127 , C12Q2523/113
摘要： This invention provides methods for nucleic acid analysis. A closed complex of nucleic acid template, nucleotide and polymerase can be formed during polymerase reaction, absent divalent metal ion. This is used to trap the labeled nucleotide complementary to the next template nucleotide in the closed complex. Detection of the label allows determination of the identity of this next correct nucleotide. Identification can be either in place, as part of the complex, or as the dye is eluted from the complex when the reaction cycle is completed by the addition of divalent metal ion. In this way, sequential nucleotides of a DNA can be identified, effectively determining the DNA sequence. This method can be applied to nucleic acid single molecules or to collections of identical or nearly identical sequence such as PCR products or clones. Multiple templates can be sequenced in parallel, particularly if they are immobilized on a solid support.
摘要翻译：本发明提供核酸分析方法。可以在聚合酶反应期间形成核酸模板，核苷酸和聚合酶的闭合复合物，不存在二价金属离子。这用于捕获与封闭复合物中下一个模板核苷酸互补的标记核苷酸。标签的检测允许确定下一个正确的核苷酸的身份。识别可以作为复合物的一部分就位，或当通过添加二价金属离子完成反应循环时染料从络合物中洗脱出来。以这种方式，可以鉴定DNA的连续核苷酸，有效地确定DNA序列。该方法可以应用于核酸单分子或相同或几乎相同序列的集合，例如PCR产物或克隆。多个模板可以并行测序，特别是如果它们固定在固体支持物上。

35. 发明申请

WO2004072297A2 TERMINAL-PHOSPHATE-LABELED NUCLEOTIDES AND METHODS OF USE 审中-公开
标题翻译：终端磷酸酯标记的核苷酸和使用方法
公开(公告)号：WO2004072297A2
公开(公告)日：2004-08-26
申请号：PCT/US2004/002785
申请日：2004-01-30
申请人： AMERSHAM BIOSCIENCES CORP , FULLER, Carl , KUMAR, Shiv , SOOD, Anup , NELSON, John
发明人： FULLER, Carl , KUMAR, Shiv , SOOD, Anup , NELSON, John
IPC分类号： C12Q
CPC分类号： C12Q1/6823 , C07H19/10 , C07H19/20 , C07H21/00 , C12Q1/6816 , C12Q1/6851 , C12Q2521/525
摘要： The present invention relates to improved methods of detecting a target using a labeled substrate or substrate analog. The methods comprise reacting the substrate or substrate analog in an enzyme-catalyzed reaction which produces a labeled moiety with independently detectable signal only when such substrate or substrate analog reacts. The present invention, in particular, describes methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrate for nucleic acid polymerases. The methods provided by this invention utilize a nucleoside polyphosphate, dideoxynucleoside polyphosphate, or deoxynucleoside polyphosphate analogue which has a colorimetric dye, chemiluminescent, or fluorescent moiety, a mass tag or an electrochemical tag attached to the terminal-phosphate. When a nucleic acid polymerase uses this analogue as a substrate, an enzyme-activatable label would be present on the inorganic polyphosphate by-product of phosphoryl transfer. Cleavage of the polyphosphate product of phosphoryl transfer via phosphatase leads to a detectable change in the label attached thereon. When the polymerase assay is performed in the presence of a phosphatase, there is provided a convenient method for real-time monitoring of DNA or RNA synthesis and detection of a target nucleic acid.
摘要翻译：本发明涉及使用标记的底物或底物类似物检测靶的改进方法。所述方法包括在酶催化的反应中使底物或底物类似物反应，其仅在这种底物或底物类似物反应时产生具有独立可检测信号的标记部分。本发明特别地描述了基于使用末端磷酸酯标记的核苷酸作为核酸聚合酶的底物来检测样品中核酸的方法。本发明提供的方法利用了具有比色染料，化学发光或荧光部分的核苷多磷酸，双脱氧核苷多聚磷酸酯或脱氧核苷多聚磷酸酯类似物，连接至末端磷酸酯的质量标签或电化学标记。当核酸聚合酶使用该类似物作为底物时，酶活性标记将存在于磷酸转移的无机多磷酸盐副产物上。通过磷酸酶切割磷酸转移的多磷酸盐产物导致附着在其上的标记物的可检测的变化。当在磷酸酶的存在下进行聚合酶测定时，提供了用于实时监测DNA或RNA合成和检测靶核酸的方便的方法。

36. 发明申请

WO2004072238A2 TERMINAL-PHOSPHATE-LABELED NUCLEOTIDES WITH NEW LINKERS 审中-公开
标题翻译：带有新连接器的终端磷酸酯标记的核苷酸
公开(公告)号：WO2004072238A2
公开(公告)日：2004-08-26
申请号：PCT/US2004/003284
申请日：2004-02-05
申请人： AMERSHAM BIOSCIENCES CORP , KUMAR, Shiv , MCDOUGALL, Mark , SOOD, Anup , NELSON, John , FULLER, Carl , MACKLIN, John , MITSIS, Paul
发明人： KUMAR, Shiv , MCDOUGALL, Mark , SOOD, Anup , NELSON, John , FULLER, Carl , MACKLIN, John , MITSIS, Paul
IPC分类号： C12N
CPC分类号： C07H19/10 , C07H19/20 , C07H21/00 , C12P19/34
摘要： The present invention describes methods of using terminal-phosphate-labeled nucleotides in the presence of a manganese salt to enhance their substrate properties towards various enzymes. Particularly described are methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates for nucleic acid polymerases, in the presence of a manganese salt. Further provided are manganese complexes of terminal-phosphate-labeled nucleotides as well as terminal-phosphate-labeled nucleotides with new linkers with enhanced substrate properties.
摘要翻译：本发明描述了在锰盐存在下使用末端磷酸酯标记的核苷酸以增强其对各种酶的底物性质的方法。特别描述的是在锰盐存在的情况下，基于使用末端磷酸酯标记的核苷酸作为核酸聚合酶的底物，检测样品中的核酸的方法。还提供了末端磷酸酯标记的核苷酸的锰络合物以及具有增强的底物性质的新接头的末端磷酸酯标记的核苷酸。

37. 发明申请

WO2004071155A2 SOLID PHASE SEQUENCING 审中-公开
标题翻译：固相序列
公开(公告)号：WO2004071155A2
公开(公告)日：2004-08-26
申请号：PCT/US2004003283
申请日：2004-02-05
申请人： AMERSHAM BIOSCIENCES CORP , SOOD ANUP , KUMAR SHIV , NELSON JOHN , FULLER CARL
发明人： SOOD ANUP , KUMAR SHIV , NELSON JOHN , FULLER CARL
IPC分类号： C12P19/34 , C12Q1/68 , G01R29/18
CPC分类号： C12Q1/6858 , C12Q1/6874 , C12Q2565/537 , C12Q2535/125 , C12Q2521/525 , C12Q2525/101
摘要： The present invention describes methods of sequencing a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates for nucleic acid polymerases. The methods provided by this invention utilize a nucleoside polyphosphate, dideoxynucleoside polyphosphate, or deoxynucleoside polyphosphate analogue which has a colorimetric dye, chemiluminescent, or fluorescent moiety, a mass tag or an electrochemical tag attached to the terminal-phosphate. When a nucleic acid polymerase uses this analogue as a substrate, an enzyme-activatable label would be present on the inorganic polyphosphate by-product of phosphoryl transfer. Cleavage of the polyphosphate product of phosphoryl transfer via phosphatase leads to a detectable change in the label attached thereon. In some instances the labeled polyphosphate may be detected directly via the label and provide information on the nucleic acid. When the polymerase assay is performed in the presence of a phosphatase, there is provided a convenient method for real-time monitoring of DNA or RNA synthesis and characterization of a target nucleic acid.
摘要翻译：本发明描述了基于使用末端磷酸酯标记的核苷酸作为核酸聚合酶的底物来测序样品中核酸的方法。本发明提供的方法利用具有比色染料，化学发光或荧光部分的核苷多磷酸，双脱氧核苷多磷酸酯或脱氧核苷多聚磷酸酯类似物，连接至末端磷酸酯的质量标签或电化学标签。当核酸聚合酶使用该类似物作为底物时，酶活性标记将存在于磷酸转移的无机多磷酸盐副产物上。通过磷酸酶切割磷酸转移的多磷酸盐产物导致附着在其上的标记物的可检测的变化。在一些情况下，标记的多磷酸盐可以通过标记物直接检测并提供关于核酸的信息。当在磷酸酶存在下进行聚合酶测定时，提供了用于实时监测DNA或RNA合成和靶核酸表征的方便的方法。

38. 发明申请

WO2004020602A3 TERMINAL PHOSPHATE BLOCKED NUCLEOSIDE POLYPHOSPHATES 审中-公开
标题翻译：磷酸盐封闭的核苷酸多磷酸盐
公开(公告)号：WO2004020602A3
公开(公告)日：2004-05-13
申请号：PCT/US0327284
申请日：2003-08-29
申请人： AMERSHAM BIOSCIENCES CORP
发明人： SOOD ANUP , KUMAR SHIV , FULLER CARL , NELSON JOHN
IPC分类号： C12N15/09 , C07H21/00 , C07H21/02 , C07H21/04 , C12N20060101 , C12P19/34 , C12Q1/68
CPC分类号： C12Q1/686 , C12Q2525/186 , C12Q2531/119
摘要： The present invention describes terminal phosphate blocked nucleoside polyphosphates that are stable at high temperature and their use in nucleic acid amplification and analysis. Current invention further describes charge modified terminal phosphate blocked nucleoside polyphosphates for improved incorporation and direct loading of nucleic acid sequencing reactions onto separating media.
摘要翻译：本发明描述了在高温下稳定的末端磷酸封端的核苷多磷酸盐及其在核酸扩增和分析中的用途。本发明进一步描述了电荷修饰的末端磷酸封端的核苷多磷酸酯，用于改善引入并将核酸测序反应直接加载到分离介质上。

39. 发明申请

WO2004020604A2 ALLELE SPECIFIC PRIMER EXTENSION 审中-公开
标题翻译： ALLELE专用主机扩展
公开(公告)号：WO2004020604A2
公开(公告)日：2004-03-11
申请号：PCT/US2003/027286
申请日：2003-08-29
申请人： AMERSHAM BIOSCIENCES CORP
发明人： SOOD, Anup , KUMAR, Shiv , FULLER, Carl , NELSON, John , MACKLIN, John
IPC分类号： C12N
CPC分类号： C12Q1/6869 , C12Q1/6823 , C12Q2600/156 , C12Q2535/125 , C12Q2525/125 , C12Q2521/125 , C12Q2565/301 , C12Q2525/101
摘要： A method of characterizing a nucleic acid sample is provided that includes the steps of: (a) conducting a DNA polymerase reaction that includes the reaction of a template, an allele specific primer, at least one terminal phosphate-labeled nucleotide, DNA polymerase, and optionally an enzyme having 3' ? 5' exonuclease activity when the primer is non-hydrolyzable, which reaction results in the production of labeled polyphosphate; (b) permitting the labeled polyphosphate to react with a phosphatase to produce a detectable species; (c) detecting the detectable species; and (d) characterizing the nucleic acid sample based on such detection.
摘要翻译：提供了表征核酸样品的方法，其包括以下步骤：（a）进行DNA聚合酶反应，其包括模板，等位基因特异性引物，至少一个末端磷酸盐标记的核苷酸，DNA聚合酶和任选的具有3' 5'外切核酸酶活性，当该引物是不可水解的时，该反应导致标记的多磷酸盐的产生; （b）使标记的多磷酸酯与磷酸酯酶反应产生可检测的物质; （c）检测可检测物质; 和（d）基于这种检测表征核酸样品。

40. 发明申请

WO2003089606A3 THERMOSTABLE DNA POLYMERASES AND METHODS OF MAKING SAME 审中-公开
公开(公告)号：WO2003089606A3
公开(公告)日：2003-10-30
申请号：PCT/US2003/012061
申请日：2003-04-17
申请人： AMERSHAM BIOSCIENCES CORP
发明人： FARCHAUS, Joseph, W., III
IPC分类号： C12N9/12
摘要： The present invention relates to methods and compositions for providing purified thermostable enzymes, particularly thermostable DNA polymerases, that are free of exogenous detergents. The present invention also provides methods for providing such purified thermostable DNA polymerases to assays in an active form by adding one or more detergents. The present invention further provides compositions and kits comprising purified thermostable DNA polymerases for use in a variety of applications, including amplification and sequencing of nucleic acids.

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