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    ISOLATION OF POLYMERASE-NUCLEIC ACID COMPLEXES AND LOADING ONTO SUBSTRATES
    5.
    发明申请
    ISOLATION OF POLYMERASE-NUCLEIC ACID COMPLEXES AND LOADING ONTO SUBSTRATES 审中-公开
    分离聚合酶 - 核酸复合物并加载到基底上

    公开(公告)号：WO2012129242A3

    公开(公告)日：2012-12-27

    申请号：PCT/US2012029830

    申请日：2012-03-20

    Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  THANG PHAM ,  GRAY JEREMY ,  FEHR ADRIAN ,  BHAMIDIPATI ARUNASHREE ,  CHMURA AJ ,  TRAVERS KEVIN ,  OLIVARES ERIC

    Inventor： THANG PHAM ,  GRAY JEREMY ,  FEHR ADRIAN ,  BHAMIDIPATI ARUNASHREE ,  CHMURA AJ ,  TRAVERS KEVIN ,  OLIVARES ERIC

    IPC: C12Q1/48 ,  C12N15/11 ,  C12Q1/68 ,  G01N27/26 ,  G01N33/68

    CPC classification number: C12Q1/6874 ,  B01L9/523 ,  C12Q1/6869 ,  C40B60/14 ,  G01N33/54313 ,  G01N33/54353 ,  G01N2333/922 ,  C12Q2521/543 ,  C12Q2523/303 ,  C12Q2525/301

    Abstract: Compositions, methods and systems are provided for the isolation of polymerase-nucleic acid complexes. Complexes can be separated from free enzyme by using hook molecules to target single stranded regions on the nucleic acid. Active complexes can be isolated from mixtures having both active and inactive complexes by initiating nucleic acid synthesis so as to open up a portion of a double stranded region rendering that region single stranded. Hook molecules are targeted to bind the sequences that are thus exposed. The hook molecules bound to active polymerase-nucleic acid complex are isolated, and the active polymerase-nucleic acid complexes released. Also disclosed are compositions, devices, and methods for loading molecules-of-interest onto a substrate by contacting beads having molecules-of-interest attached to them with the substrate, for example by providing a field that brings the beads into proximity or contact with the substrate and moves the beads with respect to the substrate. Such molecules-of-interest can be deposited onto substrates for single-molecule analysis.

    Abstract translation: 提供了用于分离聚合酶 - 核酸复合物的组合物，方法和系统。 通过使用钩分子将复合物与游离酶分离以靶向核酸上的单链区域。 活性配合物可以通过引发核酸合成从具有活性和非活性复合物的混合物中分离出来，以便打开一部分双链区域，使该区域成为单链。 钩状分子被靶向以结合由此暴露的序列。 分离与活性聚合酶 - 核酸复合物结合的钩分子，释放活性聚合酶 - 核酸复合物。 还公开了通过使具有附着于其上的感兴趣分子的珠粒与底物接触将负载分子加载到底物上的组合物，装置和方法，例如通过提供使珠子接近或接触 所述基板并相对于所述基板移动所述珠。 这种感兴趣的分子可以沉积在底物上用于单分子分析。

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