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    Detection method for gene expression
    7.
    发明授权
    Detection method for gene expression 有权
    基因表达检测方法

    公开(公告)号：US07914983B2

    公开(公告)日：2011-03-29

    申请号：US11427395

    申请日：2006-06-29

    申请人： Chockalingam Palaniappan ,  Carl W. Fuller ,  John R. Nelson

    发明人： Chockalingam Palaniappan ,  Carl W. Fuller ,  John R. Nelson

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C12M1/34 ,  C07H21/04

    CPC分类号： C12Q1/6809 ,  C12Q2565/501 ,  C12Q2521/131 ,  C12Q2563/155 ,  C12Q2535/101

    摘要： Provided is a novel approach for generating oligonucleotide probes and the use of these probes in gene expression profiling, by hybridization to test oligonucleotides on arrays or beads. This approach involves labeling of the complement oligonucleotide probes using a mixture of dye or hapten labeled-ddNTPs in solution. The labeled oligonucleotide probes are then used to hybridize to the test oligonucleotides on the solid support. Success in hybridization is monitored by associated signal on the solid support. This approach greatly reduces hybridization time, due to the simplification of the probe content. It is especially useful when analyzing a small number of genes, such as a signature set of genes for a disease or condition.

    摘要翻译： 提供了一种用于产生寡核苷酸探针的新方法，以及这些探针在基因表达谱中的应用，通过与测试寡核苷酸与阵列或珠粒杂交。 该方法包括使用染料或半抗原标记的ddNTP在溶液中的混合物来标记补体寡核苷酸探针。 然后将标记的寡核苷酸探针与固体支持物上的测试寡核苷酸杂交。 通过固相支持物上的相关信号监测杂交成功。 由于探针含量的简化，这种方法大大减少了杂交时间。 当分析少量基因（例如疾病或病症的基因的签名集）时，它是特别有用的。

    Method for nucleic acid analysis
    8.
    发明授权
    Method for nucleic acid analysis 有权
    核酸分析方法

    公开(公告)号：US07871771B2

    公开(公告)日：2011-01-18

    申请号：US11570347

    申请日：2005-06-09

    申请人： Carl W. Fuller ,  John R. Nelson

    发明人： Carl W. Fuller ,  John R. Nelson

    IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H21/04

    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。

    DNA cycle sequencing
    9.
    发明授权
    DNA cycle sequencing 失效
    DNA循环测序

    公开(公告)号：US5741676A

    公开(公告)日：1998-04-21

    申请号：US437318

    申请日：1995-05-09

    申请人： Carl W. Fuller

    发明人： Carl W. Fuller

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6806 ,  C12Q1/6869

    摘要： Amplified DNA is prepared for sequencing by contacting with exonuclease I and alkaline phosphatase which degrade undesirable excess primers dNTPs and non-specifically amplified single stranded DNA therein. The enzymes are provided in a kit. The method for sequencing DNA includes the following steps: providing a polynucleotide primer complementary to a region of the DNA, providing the DNA to be sequenced, and contacting that primer and DNA together in the presence of a DNA polymerase and between 1 and 3 dNTPs, at least one of the dNTP being labelled. The primer and DNA are contacted under conditions which allow extension of the primer by addition of one or more of the dNTPs to the primer to form an extended primer. The primer and DNA are then dissociated, generally by heating, and the contacting and dissociating steps repeated a plurality of times (usually 10-200 times). Finally, the extended primer is contacted with the DNA in the presence of a DNA polymerase (which is generally the same polymerase as used in the initial labelling step) all four dNTPs and a chain terminating agent.

    摘要翻译： 制备扩增的DNA用于通过与外切核酸酶I和碱性磷酸酶接触进行测序，其将不期望的过量引物dNTPs和非特异性扩增的单链DNA降解。 酶在试剂盒中提供。 用于测序DNA的方法包括以下步骤：提供与DNA区域互补的多核苷酸引物，提供要测序的DNA，并在DNA聚合酶和1至3 dNTP之间将该引物和DNA接触在一起， dNTP中的至少一个被标记。 引物和DNA在允许通过向引物中加入一种或多种dNTP扩增引物以形成延伸引物的条件下接触。 然后通常通过加热解离引物和DNA，并且接触和解离步骤重复多次（通常为10-200次）。 最后，将延伸的引物与DNA在所有四种dNTP和链终止剂的DNA聚合酶（其通常与初始标记步骤中使用的聚合酶通常相同）存在下接触。

    DNA cycle sequencing
    10.
    发明授权
    DNA cycle sequencing 失效
    DNA循环测序

    公开(公告)号：US5674679A

    公开(公告)日：1997-10-07

    申请号：US767137

    申请日：1991-09-27

    申请人： Carl W. Fuller

    发明人： Carl W. Fuller

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/6806 ,  C12Q1/6869

    摘要： Method for sequencing DNA which includes the following steps: providing a polynucleotide primer complementary to a region of the DNA, providing the DNA to be sequenced, and contacting that primer and DNA together in the presence of a DNA polymerase and between 1 and 3 dNTPs, at least one of the dNTP being labelled. The primer and DNA are contacted under conditions which allow extension of the primer by addition of one or more of the dNTPs to the primer to form an extended primer. The primer and DNA are then dissociated, generally by heating, and the contacting and dissociating steps repeated a plurality of times (usually 10-200 times). Finally, the extended primer is contacted with the DNA in the presence of a DNA polymerase (which is generally the same polymerase as used in the initial labelling step) all four dNTPs and a chain terminating agent.

    摘要翻译： 测序DNA的方法，其包括以下步骤：提供与DNA区域互补的多核苷酸引物，提供要测序的DNA，并在DNA聚合酶和1至3 dNTPs存在下将该引物和DNA接触， dNTP中的至少一个被标记。 引物和DNA在允许通过向引物中加入一种或多种dNTP扩增引物以形成延伸引物的条件下接触。 然后通常通过加热解离引物和DNA，并且接触和解离步骤重复多次（通常为10-200次）。 最后，将延伸的引物与DNA在所有四种dNTP和链终止剂的DNA聚合酶（其通常与初始标记步骤中使用的聚合酶通常相同）存在下接触。