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    METHODS AND COMPOSITIONS FOR LABELING NUCLEIC ACIDS
    4.
    发明申请
    METHODS AND COMPOSITIONS FOR LABELING NUCLEIC ACIDS 有权
    标记核酸的方法和组合物

    公开(公告)号：US20100311063A1

    公开(公告)日：2010-12-09

    申请号：US12785999

    申请日：2010-05-24

    Applicant: Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    Inventor： Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    IPC: C12Q1/68

    CPC classification number: C12Q1/6806 ,  C12Q1/68 ,  C12Q1/6816 ,  C12Q1/6841 ,  G01N33/5011 ,  G01N33/582 ,  C12Q2525/101 ,  C12Q2563/107

    Abstract: The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.

    Abstract translation: 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。

    Methods and compositions for labeling nucleic acids
    5.
    发明授权
    Methods and compositions for labeling nucleic acids 有权
    用于标记核酸的方法和组合物

    公开(公告)号：US07767421B2

    公开(公告)日：2010-08-03

    申请号：US11588697

    申请日：2006-10-27

    Applicant: Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    Inventor： Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    IPC: C12P19/34 ,  C07H21/02

    CPC classification number: C12Q1/6806 ,  C12Q1/68 ,  C12Q1/6816 ,  C12Q1/6841 ,  G01N33/5011 ,  G01N33/582 ,  C12Q2525/101 ,  C12Q2563/107

    Abstract: The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.

    Abstract translation: 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。

    Methods and compositions for labeling nucleic acids
    6.
    发明授权
    Methods and compositions for labeling nucleic acids 有权
    用于标记核酸的方法和组合物

    公开(公告)号：US08541570B2

    公开(公告)日：2013-09-24

    申请号：US12785999

    申请日：2010-05-24

    Applicant: Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    Inventor： Kyle R. Gee ,  Brian Agnew ,  Adrian Salic ,  Timothy J. Mitchison

    IPC: C07H19/06 ,  C07F9/02 ,  G01N33/48

    CPC classification number: C12Q1/6806 ,  C12Q1/68 ,  C12Q1/6816 ,  C12Q1/6841 ,  G01N33/5011 ,  G01N33/582 ,  C12Q2525/101 ,  C12Q2563/107

    Abstract: The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.

    Abstract translation: 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。

    Compositions and methods for detection and isolation of phosphorylated molecules
    9.
    发明授权
    Compositions and methods for detection and isolation of phosphorylated molecules 有权
    用于磷酸化分子检测和分离的组合物和方法

    公开(公告)号：US07445894B2

    公开(公告)日：2008-11-04

    申请号：US10821522

    申请日：2004-04-09

    Applicant: Brian Agnew ,  Kyle R. Gee ,  Vladimir V. Martin

    Inventor： Brian Agnew ,  Kyle R. Gee ,  Vladimir V. Martin

    IPC: G01N33/50

    CPC classification number: G01N33/5008 ,  G01N33/502 ,  G01N33/5091 ,  G01N2333/4709

    Abstract: The present invention relates to phosphate-binding compounds that find use in binding, detecting and isolating phosphorylated target molecules including the subsequent identification of target molecules that interact with phosphorylated target molecules or molecules capable of being phosphorylated. A binding solution is provide that comprises a phosphate-binding compound, an acid and a metal ion wherein the metal ion simultaneously interacts with an exposed phosphate group on a target molecule and the metal chelating moiety of the phosphate-binding compound forming a bridge between the phosphate-binding compound and a phosphorylated target molecule resulting in a ternary complex. The binding solution of the present invention finds use in binding and detecting immobilized and solubilized phosphorylated target molecules, isolation of phosphorylated target molecules from a complex mixture and aiding in proteomic analysis wherein kinase and phosphatase substrates and enzymes can be identified.

    Abstract translation: 本发明涉及用于结合，检测和分离磷酸化靶分子的磷酸盐结合化合物，包括随后鉴定与磷酸化靶分子或能够被磷酸化的分子相互作用的靶分子。 提供的结合溶液包括磷酸盐结合化合物，酸和金属离子，其中金属离子同时与靶分子上暴露的磷酸基团相互作用，并且磷酸盐结合化合物的金属螯合部分形成在 磷酸化结合化合物和导致三元复合物的磷酸化靶分子。 本发明的结合溶液用于结合和检测固定和溶解的磷酸化靶分子，从复杂混合物中分离磷酸化靶分子，并辅助蛋白质组分析，其中可以鉴定激酶和磷酸酶底物和酶。

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