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    Polymerase compositions, methods of making and using same
    1.
    发明授权
    Polymerase compositions, methods of making and using same 有权

    公开(公告)号：US11821011B2

    公开(公告)日：2023-11-21

    申请号：US17445047

    申请日：2021-08-13

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Peter Vander Horn ,  Daniel Mazur ,  Theo Nikiforov ,  Mindy Landes ,  Eileen Tozer

    IPC: C12N9/12 ,  C12Q1/68 ,  C12Q1/686 ,  C12Q1/6869

    CPC classification number: C12N9/1252 ,  C12Q1/686 ,  C12Q1/6869

    Abstract: The present disclosure provides compositions, methods, kits, systems and apparatus that are useful for nucleic acid polymerization. In particular, modified polymerases and biologically active fragments thereof are provided that allow for nucleic acid amplification. In some aspects, the disclosure provides modified polymerases having lower systematic error as compared to a reference polymerase. In one aspect, the disclosure relates to modified polymerases useful for nucleic acid sequencing, genotyping, copy number variation analysis, paired-end sequencing and other forms of genetic analysis. In some aspects, the disclosure relates to modified polymerases useful for the generation of nucleic acid libraries or nucleic acid templates. In some aspects, the disclosure relates to the identification of homologous amino acid mutations that can be transferred across classes or families of polymerases to provide novel polymerases with altered properties.

    Labeled enzyme compositions, methods and systems
    2.
    发明授权
    Labeled enzyme compositions, methods and systems 有权

    公开(公告)号：US11542549B2

    公开(公告)日：2023-01-03

    申请号：US15907845

    申请日：2018-02-28

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Theo Nikiforov

    IPC: C12N9/12 ,  C12Q1/6869 ,  C07H19/20 ,  C12N9/96 ,  C12Q1/6818 ,  G01N33/58 ,  G01N21/64

    Abstract: Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are labeled polymerase conjugates including a polymerase linked to one or more labels, wherein the conjugate has polymerase activity. Such conjugates can exhibit enhanced biological activity and/or superior detectability as compared to conventional labeled polymerases. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.

    Polymerase compositions, methods of making and using same
    4.
    发明授权
    Polymerase compositions, methods of making and using same 有权

    公开(公告)号：US10240134B2

    公开(公告)日：2019-03-26

    申请号：US15481858

    申请日：2017-04-07

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Peter Vander Horn ,  Daniel Mazur ,  Theo Nikiforov ,  Mindy Landes ,  Eileen Tozer

    IPC: C12N9/12 ,  C12Q1/68 ,  C12Q1/686 ,  C12Q1/6869

    Abstract: The present disclosure provides compositions, methods, kits, systems and apparatus that are useful for nucleic acid polymerization. In particular, modified polymerases and biologically active fragments thereof are provided that allow for nucleic acid amplification. In some aspects, the disclosure provides modified polymerases having lower systematic error as compared to a reference polymerase. In one aspect, the disclosure relates to modified polymerases useful for nucleic acid sequencing, genotyping, copy number variation analysis, paired-end sequencing and other forms of genetic analysis. In some aspects, the disclosure relates to modified polymerases useful for the generation of nucleic acid libraries or nucleic acid templates. In some aspects, the disclosure relates to the identification of homologous amino acid mutations that can be transferred across classes or families of polymerases to provide novel polymerases with altered properties.

    Methods and compositions for nucleic acid amplification
    6.
    发明授权
    Methods and compositions for nucleic acid amplification 有权

    公开(公告)号：US11268117B2

    公开(公告)日：2022-03-08

    申请号：US16308803

    申请日：2017-06-09

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Hua Yu ,  Theo Nikiforov ,  Abraham Rosenbaum

    IPC: C12P19/34 ,  C12Q1/6853 ,  C12Q1/6844

    Abstract: In some embodiments, the disclosure relates generally to methods, as well as related compositions and kits for recombinase-mediated nucleic acid amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using at least one blocked primer that contains a 5′ domain, at least one nucleotide that is cleavable by an RNase H enzyme, a 3′ domain, wherein the primer is not extendable by a polymerase, and wherein the 3′ domain has a length of 7-100 nucleotides, for example 10-30 nucleotides. These methods and the use of a blocked primer reduce or eliminate non-specific amplification products, such as primer dimers, which are generated in RPA reactions.

    Thiolated nucleotide analogues for nucleic acid synthesis
    7.
    发明授权
    Thiolated nucleotide analogues for nucleic acid synthesis 有权

    公开(公告)号：US10344325B2

    公开(公告)日：2019-07-09

    申请号：US14937211

    申请日：2015-11-10

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Steven Menchen ,  Theo Nikiforov ,  Barnett Rosenblum

    IPC: C12Q1/68 ,  C12Q1/6869 ,  G01N27/414

    Abstract: The present disclosure provide systems, compositions, methods, reagents, kits and products for extending a nucleic acid that includes incorporating a nucleotide residue at a terminus of a nucleic acid using a polymerase enzyme and at least one nucleotide, wherein the at least one nucleotide includes a thiophosphate moiety, and wherein the at least one nucleotide is resistant to hydrolysis by phosphatase. In some embodiments, the nucleotide incorporation can be conducted in the presence of a phosphatase. In some embodiments, the nucleotide incorporation can be conducted in the presence of at least on chelation moiety that is configured to bind an orthophosphate moiety.

    Bi-directional sequencing compositions and methods
    9.
    发明授权
    Bi-directional sequencing compositions and methods 有权
    双向测序组合物和方法

    公开(公告)号：US09587269B2

    公开(公告)日：2017-03-07

    申请号：US14832772

    申请日：2015-08-21

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Jason Myers ,  Zhoutao Chen ,  Devin Dressman ,  Theo Nikiforov

    IPC: C12P19/34 ,  C12Q1/68

    CPC classification number: C12Q1/6874 ,  C12Q1/6806 ,  C12Q1/6869 ,  C12Q2521/307 ,  C12Q2521/325 ,  C12Q2525/125 ,  C12Q2535/119 ,  C12Q2565/518

    Abstract: In some embodiments, methods for obtaining sequence information from a nucleic acid template linked to a support include hybridizing a first primer to a template strand linked to a support, sequencing a portion of the nucleic acid template, thereby forming an extended first primer product that is complementary to a portion of the nucleic acid template, In some embodiments, the method further includes introducing a nick into a portion of the template strand that is hybridized to the extended first primer product, degrading a portion of the template strand from the nick using a degrading agent, where a portion of the extended first primer remains hybridized to an undegraded portion of the template strand, and sequencing at least some of the single-stranded portion of the extended first primer by synthesis.

    Abstract translation: 在一些实施方案中，从与支持物连接的核酸模板获得序列信息的方法包括将第一引物与连接于载体的模板链杂交，对核酸模板的一部分进行测序，从而形成延伸的第一引物产物， 与一部分核酸模板互补。在一些实施方案中，该方法还包括将切口引入与延伸的第一引物产物杂交的模板链的一部分中，使用下述方法从模板中降解模板链的一部分： 其中一部分延伸的第一引物保留与模板链的未降解部分杂交，并通过合成测序延伸的第一引物的至少一些单链部分。

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