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    Immobilized nucleic acid complexes for sequence analysis
    31.
    发明授权
    Immobilized nucleic acid complexes for sequence analysis 有权
    固定核酸复合物进行序列分析

    公开(公告)号：US08481264B2

    公开(公告)日：2013-07-09

    申请号：US12562690

    申请日：2009-09-18

    Applicant: Keith Bjornson ,  Arek Bibillo ,  Fred Christians ,  Kevin Travers ,  Robin Emig

    Inventor： Keith Bjornson ,  Arek Bibillo ,  Fred Christians ,  Kevin Travers ,  Robin Emig

    IPC: C12Q1/68 ,  C12P19/34 ,  C07H21/04

    CPC classification number: C12Q1/6874 ,  C12Q2531/125

    Abstract: Provided are methods for sequencing a nucleic acid that include fixing a template to a surface through a template localizing moiety and sequencing the nucleic acid with a sequencing enzyme, e.g. a polymerase or exonuclease. The sequencing enzyme can optionally be exchanged with a second sequencing enzyme, which continues the sequencing of the nucleic acid. The template localizing moiety can optionally anneal with the nucleic acid and/or associate with the sequencing enzyme. Also provided are compositions comprising a nucleic acid fixed to a surface via a template localizing moiety, and a first sequencing enzyme, which can sequence the nucleic acid and optionally exchange with a second sequencing enzyme present in the composition. Compositions in which a template localizing moiety is immobilized on a surface are provided. Compositions for sequencing reactions are provided. Also provided are sequencing systems comprising reaction regions in which or near which template localizing moieties are immobilized.

    Abstract translation: 提供了用于对核酸进行测序的方法，其包括通过模板定位部分将模板固定到表面，并用测序酶（例如）测序核酸。 聚合酶或外切核酸酶。 测序酶可以任选地与第二个测序酶交换，其继续测序核酸。 模板定位部分可任选地与核酸退火和/或与测序酶缔合。 还提供了包含通过模板定位部分固定在表面上的核酸的组合物和可以使序列核酸并任选地与组合物中存在的第二测序酶交换的第一测序酶。 提供其中将模板定位部分固定在表面上的组合物。 提供了测序反应的组合物。 还提供了包含反应区的测序系统，其中固定有定位部分的模板定位部分。

    Generation of modified polymerases for improved accuracy in single molecule sequencing
    32.
    发明授权
    Generation of modified polymerases for improved accuracy in single molecule sequencing 有权
    修饰聚合酶的生成，提高了单分子测序的准确性

    公开(公告)号：US08420366B2

    公开(公告)日：2013-04-16

    申请号：US12924701

    申请日：2010-09-30

    Applicant: Sonya Clark ,  Arek Bibillo ,  Paul Peluso ,  Fred Christians ,  Molly He ,  Insil Park ,  Harold Lee ,  Keith Bjornson ,  Lei Jia ,  Robin Emig

    Inventor： Sonya Clark ,  Arek Bibillo ,  Paul Peluso ,  Fred Christians ,  Molly He ,  Insil Park ,  Harold Lee ,  Keith Bjornson ,  Lei Jia ,  Robin Emig

    IPC: C12N9/12 ,  C12P19/34

    CPC classification number: C12N9/12 ,  C12N11/00 ,  C12P19/34 ,  C12Q1/68

    Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions and/or heterologous or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include reduced reaction rates at one or more steps of the polymerase kinetic cycle, increased closed polymerase/DNA complex stability, enhanced metal ion coordination, reduced exonuclease activity, decreased branching fractions, and the like. Polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog, are also provided. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

    Abstract translation: 提供了包含重组DNA聚合酶的组合物，其包括氨基酸取代，插入，缺失和/或异源或外源特征，其在聚合酶上赋予改进的性质以用于增强的单分子测序。 这种性质可以包括聚合酶动力学循环的一个或多个步骤的降低的反应速率，增加的闭合聚合酶/ DNA复合物稳定性，增强的金属离子配位，减少的核酸外切酶活性，降低的支化部分等。 还提供了表现出低于其衍生的聚合酶的支化级分的支化级分的聚合酶，或者对于磷酸酯标记的核苷酸类似物的支化级分小于约25％。 还提供了编码具有上述表型的聚合酶的核酸，以及使用这种聚合酶制备DNA或顺序DNA模板的方法。

    SINGLE MOLECULE SEQUENCING WITH TWO DISTINCT CHEMISTRY STEPS
    34.
    发明申请
    SINGLE MOLECULE SEQUENCING WITH TWO DISTINCT CHEMISTRY STEPS 有权
    单分子序列与两个不同的化学步骤

    公开(公告)号：US20110212437A1

    公开(公告)日：2011-09-01

    申请号：US13022926

    申请日：2011-02-08

    Applicant: Robin Emig ,  Lei Jia ,  Jeremiah Hanes ,  Lubomir Sebo

    Inventor： Robin Emig ,  Lei Jia ,  Jeremiah Hanes ,  Lubomir Sebo

    IPC: C12Q1/68 ,  C12M1/34 ,  C07H19/04

    CPC classification number: C12Q1/6869 ,  C07H19/10 ,  C07H19/11 ,  C07H19/20 ,  C07H19/213 ,  C12Q1/6874 ,  C12Q2521/301 ,  C12Q2525/186 ,  C12Q2563/107 ,  C12Q2537/157 ,  C12Q2521/319 ,  C12Q2521/525

    Abstract: Methods, Compositions, and Systems are provided for nucleic acid sequencing where the sequential incorporation of nucleotides uses two distinct chemical steps. A plurality of nucleotide analogs, each having a labeled leaving group at its 3′ hydroxyl can be sequentially added to a growing strand in the presence of a selective cleaving activity that cleaves the 3′ hydroxyl leaving group preferentially after it has been incorporated. The selective cleaving agent can comprise an exonuclease activity, and the exonuclease activity can be a polymerase-associated exonuclease activity. Nucleotide analogs having labels on both a cleavable polyphosphate portion and on a 3′ hydroxyl leaving group can provide signals characteristic of nucleotide analog incorporation. Systems having illumination optics, collection optics, and substrates observe signals from the labels as they are being incorporated into a growing nucleic acid strand, allowing for the sequencing of template nucleic acids.

    Abstract translation: 提供了用于核酸测序的方法，组合物和系统，其中连续掺入核苷酸使用两个不同的化学步骤。 在选择性切割活性的存在下，可以将多个在其3'羟基上具有标记的离去基团的核苷酸类似物依次加入到生长链中，该选择性裂解活性优选在其引入后优先切割3'羟基离去基团。 选择性切割剂可以包含核酸外切酶活性，并且核酸外切酶活性可以是聚合酶相关的外切核酸酶活性。 在可切割聚磷酸酯部分和3'羟基离去基团上具有标记的核苷酸类似物可以提供核苷酸类似物掺入特征的信号。 具有照明光学器件，收集光学元件和底物的系统在标记被并入生长的核酸链中观察信号，允许对模板核酸进行测序。

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