公开(公告)号：US12065643B2

公开(公告)日：2024-08-20

申请号：US17302674

申请日：2021-05-10

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Mark Andersen ,  Daniel Mazur ,  Sihong Chen ,  Guobin Luo ,  Xinzhan Peng

IPC: C12N15/10 ,  C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q1/686 ,  C40B40/06 ,  C40B20/04 ,  C40B50/06

CPC classification number: C12N15/1068 ,  C12N15/1065 ,  C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q1/686 ,  C40B20/04 ,  C40B40/06 ,  C40B50/06 ,  C12Q1/6806 ,  C12Q2521/531 ,  C12Q2525/119 ,  C12Q2525/121 ,  C12Q2525/131 ,  C12Q2525/155 ,  C12Q2525/179 ,  C12Q2531/113 ,  C12Q2535/122 ,  C12Q2537/143 ,  C12Q2549/119 ,  C12Q2563/179 ,  C12Q1/6853 ,  C12Q2521/531 ,  C12Q2525/119 ,  C12Q2525/121 ,  C12Q2525/131 ,  C12Q2525/155 ,  C12Q2525/179 ,  C12Q2531/113 ,  C12Q2535/122 ,  C12Q2537/143 ,  C12Q2549/119 ,  C12Q2563/179

Abstract: Provided are methods for preparing a library of target nucleic acid sequences, as well as compositions and uses therefor. Methods comprise contacting a nucleic acid sample with a plurality of adaptors capable of amplification of one or more target nucleic acid sequences under conditions wherein the target nucleic acid(s) undergo a first amplification; digesting the resulting first amplification products; repairing the digested target amplicons; and amplifying the repaired products in a second amplification, thereby producing a library of target nucleic acid sequence. Each of the plurality of adaptor compositions comprise a handle and a targeted nucleic acid sequence and optionally one or more tag sequences. Provided methods may be carried out in a single, addition only workflow reaction, allowing for rapid production of highly multiplexed targeted libraries, optionally including unique tag sequences. Resulting library compositions are useful for a variety of applications, including sequencing applications.

公开(公告)号：US09365839B2

公开(公告)日：2016-06-14

申请号：US14710160

申请日：2015-05-12

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Stephen P. Hendricks ,  Michael Phelan ,  Marian Peris ,  Cheng-Yao Chen ,  Daniel Mazur ,  Xinzhan Peng ,  Amy Castillo

IPC: C12N9/12 ,  C07H19/20 ,  C12Q1/68

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

Abstract translation: 本文公开了显示改变的聚合酶活性的修饰聚合酶组合物，其可用于多种生物应用。 本文还公开了制备和使用这些组合物的方法。 在一些实施方案中，组合物显示改变的性质，其可以增强其在各种生物应用中的效用。 这种改变的性质可以包括例如改变的核苷酸结合亲和力，改变的核苷酸掺入动力学，改变的光稳定性和/或改变的纳米颗粒耐受性，以及本文公开的一系列其它性质。

公开(公告)号：US10093974B2

公开(公告)日：2018-10-09

申请号：US15621341

申请日：2017-06-13

Applicant: Life Technologies Corporation

Inventor： Joseph Beechem ,  Theo Nikiforov ,  Vi-En Choong ,  Xinzhan Peng ,  Guobin Luo ,  Cheng-Yao Chen ,  Michael Previte

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

Abstract: Provided herein are systems and methods for nucleotide incorporation reactions. The systems comprise polymerases having altered nucleotide incorporation kinetics and are linked to an energy transfer donor moiety, and nucleotide molecules linked with at least one energy transfer acceptor moiety. The donor and acceptor moieties undergo energy transfer when the polymerase and nucleotide are proximal to each other during nucleotide binding and/or nucleotide incorporation. As the donor and acceptor moieties undergo energy transfer, they generate an energy transfer signal which can be associated with nucleotide binding or incorporation. Detecting a time sequence of the generated signals, or the change in the signals, can be used to determine the order of the incorporated nucleotides, and can therefore be used to deduce the sequence of the target molecule.

公开(公告)号：US10093973B2

公开(公告)日：2018-10-09

申请号：US15169854

申请日：2016-06-01

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Stephen P. Hendricks ,  Michael Phelan ,  Marian Peris ,  Cheng-Yao Chen ,  Daniel Mazur ,  Xinzhan Peng ,  Amy Castillo

IPC: C12P19/34 ,  C12Q1/6869 ,  C07H19/20 ,  C12N9/12 ,  C12N9/96 ,  C12Q1/6818 ,  G01N33/58 ,  G01N21/64

Abstract: Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

公开(公告)号：US20230055565A1

公开(公告)日：2023-02-23

申请号：US17935406

申请日：2022-09-26

Applicant: Life Technologies Corporation

Inventor： Stephen P. Hendricks ,  Michael Phelan ,  Marian Peris ,  Cheng-Yao Chen ,  Daniel Mazur ,  Xinzhan Peng ,  Amy Castillo

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

Abstract: Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

公开(公告)号：US11015220B2

公开(公告)日：2021-05-25

申请号：US16123166

申请日：2018-09-06

Applicant: Life Technologies Corporation

Inventor： Theo Nikiforov ,  Daniel Mazur ,  Xinzhan Peng ,  Tommie Lloyd Lincecum ,  Yuri Belosludtsev ,  Howard Reese ,  Dmitriy Gremyachinskiy ,  Roman Rozhkov ,  John Mauro ,  Joseph Beechem ,  Eric Tulsky ,  Imad Naasani ,  Kari Haley ,  Joseph Treadway

IPC: C12Q1/6869 ,  C07H19/20 ,  C12N9/12 ,  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 polymerase-nanoparticle conjugates including a polymerase linked to a nanoparticle, wherein the conjugate has polymerase activity. Such conjugates can exhibit reduced aggregation and improved stochiometries wherein the average biomolecule:nanoparticle ratio approaches or equals 1:1. 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.

公开(公告)号：US11001836B2

公开(公告)日：2021-05-11

申请号：US16024175

申请日：2018-06-29

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Mark Andersen ,  Daniel Mazur ,  Sihong Chen ,  Guobin Luo ,  Xinzhan Peng

IPC: C12N15/10 ,  C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q1/686 ,  C40B20/04 ,  C40B50/06 ,  C40B40/06

Abstract: Provided are methods for preparing a library of target nucleic acid sequences, as well as compositions and uses therefor. Methods comprise contacting a nucleic acid sample with a plurality of adaptors capable of amplification of one or more target nucleic acid sequences under conditions wherein the target nucleic acid(s) undergo a first amplification; digesting the resulting first amplification products; repairing the digested target amplicons; and amplifying the repaired products in a second amplification, thereby producing a library of target nucleic acid sequence. Each of the plurality of adaptor compositions comprise a handle and a targeted nucleic acid sequence and optionally one or more tag sequences. Provided methods may be carried out in a single, addition only workflow reaction, allowing for rapid production of highly multiplexed targeted libraries, optionally including unique tag sequences. Resulting library compositions are useful for a variety of applications, including sequencing applications.

公开(公告)号：US12163188B2

公开(公告)日：2024-12-10

申请号：US17935406

申请日：2022-09-26

Applicant: Life Technologies Corporation

Inventor： Stephen P. Hendricks ,  Michael Phelan ,  Marian Peris ,  Cheng-Yao Chen ,  Daniel Mazur ,  Xinzhan Peng ,  Amy Castillo

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

Abstract: Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

公开(公告)号：US11453909B2

公开(公告)日：2022-09-27

申请号：US16123104

申请日：2018-09-06

Applicant: Life Technologies Corporation

Inventor： Stephen P. Hendricks ,  Michael Phelan ,  Marian Peris ,  Cheng-Yao Chen ,  Daniel Mazur ,  Xinzhan Peng ,  Amy Castillo

IPC: C12N9/12 ,  C12P19/34 ,  C12Q1/6869 ,  C07H19/20 ,  C12N9/96 ,  C12Q1/6818 ,  G01N33/58 ,  G01N21/64

Abstract: Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.

公开(公告)号：US09695471B2

公开(公告)日：2017-07-04

申请号：US14584829

申请日：2014-12-29

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Joseph Beechem ,  Theo Nikiforov ,  Vi-En Choong ,  Xinzhan Peng ,  Guobin Luo ,  Cheng-Yao Chen ,  Michael Previte

IPC: C12P19/34 ,  C12N9/12 ,  C12Q1/68 ,  C07H19/20 ,  C12N9/96 ,  G01N33/58 ,  G01N21/64

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Provided herein are systems and methods for nucleotide incorporation reactions. The systems comprise polymerases having altered nucleotide incorporation kinetics and are linked to an energy transfer donor moiety, and nucleotide molecules linked with at least one energy transfer acceptor moiety. The donor and acceptor moieties undergo energy transfer when the polymerase and nucleotide are proximal to each other during nucleotide binding and/or nucleotide incorporation. As the donor and acceptor moieties undergo energy transfer, they generate an energy transfer signal which can be associated with nucleotide binding or incorporation. Detecting a time sequence of the generated signals, or the change in the signals, can be used to determine the order of the incorporated nucleotides, and can therefore be used to deduce the sequence of the target molecule.