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    Single nucleotide detection method and associated probes
    8.
    发明授权
    Single nucleotide detection method and associated probes 有权

    公开(公告)号：US11920192B2

    公开(公告)日：2024-03-05

    申请号：US16613593

    申请日：2018-05-15

    Applicant: LIGHTCAST DISCOVERY LTD

    Inventor： Barnaby Balmforth ,  Cameron Alexander Frayling ,  Mark Dethlefsen

    IPC: C12Q1/6869 ,  C12Q1/6823

    CPC classification number: C12Q1/6869 ,  C12Q1/6823 ,  C12Q2521/101 ,  C12Q2521/301 ,  C12Q2521/319 ,  C12Q2525/125 ,  C12Q2525/307 ,  C12Q2537/149 ,  C12Q2563/107 ,  C12Q2563/159

    Abstract: A method of sequencing a nucleic acid comprising (1) generating a stream of single nucleoside triphosphates by progressive enzymatic digestion of the nucleic acid; (2) producing at least one oligonucleotide used probe by reacting, in the presence of a polymerase, at least one of the single nucleoside triphosphates with a corresponding biological probe comprising (a) a first single-stranded oligonucleotide including an exonuclease blocking-site, a restriction endonuclease recognition-site located on the 5′ side of the blocking-site and including a single nucleotide capture-site e, and at least one fluorophore region and (b) a second and optionally a third single-stranded oligonucleotide each separate from the first oligonucleotide; (3) cleaving the first oligonucleotide strand of the used probe at the recognition-site with a restriction endonuclease; (4) digesting the first oligonucleotide component with an enzyme to yield fluorophores in a detectable state and (5) detecting the fluorophores released in step (4).

    AMPLIFICATION OF NUCLEIC ACIDS
    9.
    发明公开
    AMPLIFICATION OF NUCLEIC ACIDS 审中-公开

    公开(公告)号：US20240026433A1

    公开(公告)日：2024-01-25

    申请号：US18304282

    申请日：2023-04-20

    Applicant: LumiraDx UK Ltd.

    Inventor： Daiwei Shen ,  Bryan Kraynack ,  Victor Perez ,  Jarrod Provins

    IPC: C12Q1/6848 ,  C12Q1/6851 ,  C12Q1/6853

    CPC classification number: C12Q1/6848 ,  C12Q1/6851 ,  C12Q1/6853 ,  C12Q2521/101 ,  C12Q2521/107 ,  C12Q2521/307 ,  C12Q2525/113 ,  C12Q2525/131 ,  C12Q2525/301 ,  C12Q2527/101 ,  C12Q2537/137

    Abstract: Disclosed is a method of performing a non-isothermal nucleic acid amplification reaction, the method comprising the steps of: (a) mixing a target sequence with one or more complementary single stranded primers in conditions which permit a hybridisation event in which the primers hybridise to the target, which hybridisation event, directly or indirectly, leads to the formation of a duplex structure comprising two nicking sites disposed at or near opposite ends of the duplex; and performing an amplification process by; (b) using a nicking enzyme to cause a nick at each of said nicking sites in the strands of the duplex; (c) using a polymerase to extend the nicked strands to as to form newly synthesised nucleic acid, which extension with the polymerase recreates nicking sites; (d) repeating steps (b) and (c) as desired so as to cause the production of multiple copies of the newly synthesised nucleic acid; characterised in that the temperature at which the method is performed is non-isothermal, and subject to shuttling, a plurality of times, between an upper temperature and a lower temperature during the amplification process of steps (b)-(d), wherein at the upper temperature, one of said polymerase or nicking enzyme is more active than the other of said enzymes, such that there is a disparity in the activity of the enzymes, and at the lower temperature the disparity in the activity of the enzymes is reduced or reversed.

    Polymerase compositions and kits, and methods of using and making the same
    10.
    发明授权
    Polymerase compositions and kits, and methods of using and making the same 有权

    公开(公告)号：US11866740B2

    公开(公告)日：2024-01-09

    申请号：US17302621

    申请日：2021-05-07

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Daniel Mazur ,  Peter B. Vander Horn ,  Eileen Tozer ,  Sihong Chen ,  Guobin Luo ,  Joshua Shirley ,  Kevin Heinemann

    IPC: C12N9/12 ,  C12Q1/686

    CPC classification number: C12N9/1252 ,  C12Q1/686 ,  C12Y207/07007 ,  C12Q1/686 ,  C12Q2521/101

    Abstract: The present disclosure provides compositions, methods, kits, systems and apparatus that are useful for nucleic acid polymerization. In particular, recombinant polymerases and biologically active fragments thereof are provided that allow for nucleic acid amplification. In some aspects, the disclosure provides recombinant polymerases that yield lower systematic error rates and/or improved accuracy, when used in sequencing by synthesis reactions as compared to a control polymerase. In one aspect, the disclosure relates to recombinant polymerases useful for nucleic acid sequencing, genotyping, copy number variation analysis, paired-end sequencing and other forms of genetic analysis. In another aspect, the recombinant polymerases are useful for the amplification of nucleic acid templates during PCR, emPCR, isothermal amplification, recombinase polymerase amplification, rolling circle amplification, strand displacement amplification and proximity ligation amplification. In some aspects, the disclosure relates to recombinant polymerases useful for the generation of nucleic acid libraries and/or nucleic acid templates.

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