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    Recombinant polymerases for improved single molecule sequencing
    1.
    发明授权
    Recombinant polymerases for improved single molecule sequencing 有权
    用于改进单分子测序的重组聚合酶

    公开(公告)号：US08999676B2

    公开(公告)日：2015-04-07

    申请号：US13176697

    申请日：2011-07-05

    Applicant: Robin Emig ,  Lei Jia ,  Satwik Kamtekar ,  Erik Miller ,  Colleen Cutcliffe ,  Walter Lee

    Inventor： Robin Emig ,  Lei Jia ,  Satwik Kamtekar ,  Erik Miller ,  Colleen Cutcliffe ,  Walter Lee

    IPC: C12N15/54 ,  C12N9/00 ,  C12N9/12 ,  C12Q1/68

    CPC classification number: C12N9/1252 ,  C12N9/1241 ,  C12P19/34 ,  C12Q1/6869

    Abstract: Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. 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，其包括赋予聚合酶修饰性质以增强单分子测序的氨基酸取代，插入，缺失和/或外源特征。 这种性质可以包括增强的金属离子配位，降低的核酸外切酶活性，在聚合酶动力学循环的一个或多个步骤中降低的反应速率，降低的支化部分，改变的辅因子选择性，增加的产率，增加的热稳定性，增加的准确度，增加的速度，增加的读长度， 等等。 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
    8.
    发明授权
    Single molecule sequencing with two distinct chemistry steps 有权
    具有两个不同化学步骤的单分子测序

    公开(公告)号：US08603741B2

    公开(公告)日：2013-12-10

    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 ,  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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