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：US20170044506A1

公开(公告)日：2017-02-16

申请号：US15243808

申请日：2016-08-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Lei (Larry) XI ,  Roland Nagel ,  Stephen Hendricks ,  Jennifer Berkman ,  Marian Peris ,  Yulei Wang

IPC: C12N9/12 ,  C12N15/10

CPC classification number: C12N9/1276 ,  C12N15/1096 ,  C12Q2521/107 ,  C12Y207/07049

Abstract: The present invention provides compositions and methods for a reverse transcription reaction using a reversibly inactivated reverse transcriptase enzyme. The reversibly inactivated reverse transcriptase enzyme results from a chemical modification which inactivates the reverse transcriptase enzyme. The activity of the reverse transcriptase enzyme is recovered by an incubation of the reaction mixture at elevated temperature prior to, or as part of the reverse transcription reaction. The reverse transcriptase enzyme of the present invention provides for a significant reduction in non-specific reverse transcription from template nucleic acid molecules because the formulation of the reaction mixture does not support the formation of reverse transcription products prior to activation of the reverse transcriptase.

Abstract translation: 本发明提供使用可逆失活的逆转录酶进行逆转录反应的组合物和方法。 可逆灭活的逆转录酶由化学修饰产生，其使逆转录酶失活。 通过在反转录反应之前或作为逆转录反应的一部分的升高温度下温育反应物来回收逆转录酶的活性。 本发明的逆转录酶提供了来自模板核酸分子的非特异性逆转录的显着降低，因为反应混合物的配方不支持在逆转录酶激活之前形成逆转录产物。

公开(公告)号：US10597642B2

公开(公告)日：2020-03-24

申请号：US15690945

申请日：2017-08-30

Applicant: Life Technologies Corporation

Inventor： Peter Vander Horn ,  Cheng-Yao Chen ,  Guobin Luo ,  Michael Previte ,  Jamshid Temirov ,  Theo Nikiforov ,  Zhaohui Zhou ,  Hongye Sun ,  Yufang Wang ,  Stefanie Yukiko Nishimura ,  Hongyi Wang ,  Marian Peris ,  Barnett Rosenblum ,  Michael Phelan

IPC: C12Q1/68 ,  C12N9/12 ,  C12Q1/6827 ,  C12Q1/6869 ,  C12Q1/6872 ,  C12Q1/6804 ,  C12Q1/6818

Abstract: Provided herein are compositions and systems for use in polymerase-dependent, nucleotide transient-binding methods. The methods are useful for deducing the sequence of a template nucleic acid molecule and single nucleotide polymorphism (SNP) analyses. The methods rely on the fact that the polymerase transient-binding time for a complementary nucleotide is longer compared to that of a non-complementary nucleotide. The labeled nucleotides transiently-binds the polymerase in a template-dependent manner, but does not incorporate. The methods are conducted under any reaction condition that permits transient binding of a complementary or non-complementary nucleotide to a polymerase, and inhibits nucleotide incorporation.

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：US09765310B2

公开(公告)日：2017-09-19

申请号：US14991230

申请日：2016-01-08

Applicant: Life Technologies Corporation

Inventor： Peter Vander Horn ,  Cheng-Yao Chen ,  Guobin Luo ,  Michael Previte ,  Jamshid Temirov ,  Theo Nikiforov ,  Zhaohui Zhou ,  Hongye Sun ,  Yufang Wang ,  Stefanie Yukiko Nishimura ,  Hongyi Wang ,  Marian Peris ,  Barnett Rosenblum ,  Michael Phelan

IPC: C12Q1/68 ,  C12N9/12

CPC classification number: C12N9/1252 ,  C12Q1/68 ,  C12Q1/6804 ,  C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6869 ,  C12Q1/6872 ,  C12Y207/07007 ,  G01N2500/00 ,  C12Q2565/632 ,  C12Q2563/137 ,  C12Q2563/107 ,  C12Q2565/30 ,  C12Q2537/157

Abstract: Provided herein are compositions and systems for use in polymerase-dependent, nucleotide transient-binding methods. The methods are useful for deducing the sequence of a template nucleic acid molecule and single nucleotide polymorphism (SNP) analyses. The methods rely on the fact that the polymerase transient-binding time for a complementary nucleotide is longer compared to that of a non-complementary nucleotide. The labeled nucleotides transiently-binds the polymerase in a template-dependent manner, but does not incorporate. The methods are conducted under any reaction condition that permits transient binding of a complementary or non-complementary nucleotide to a polymerase, and inhibits nucleotide incorporation.

公开(公告)号：US09399767B2

公开(公告)日：2016-07-26

申请号：US14191997

申请日：2014-02-27

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Marian Peris ,  Michael Phelan ,  Barnett Rosenblum ,  Stephen Hendricks

IPC: C12N9/12 ,  C12Q1/68

CPC classification number: C12N9/1252 ,  C12Q1/68 ,  C12Q1/6804 ,  C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6869 ,  C12Q1/6872 ,  C12Y207/07007 ,  G01N2500/00 ,  C12Q2565/632 ,  C12Q2563/137 ,  C12Q2563/107 ,  C12Q2565/30 ,  C12Q2537/157

Abstract: Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.

Abstract translation: 本文提供了衍生自野生型RB69 DNA聚合酶或与野生型RB69 DNA聚合酶相关的突变型DNA依赖性聚合酶。 这些突变型聚合酶能够选择性地结合标记的核苷酸。 这些突变型聚合酶还能够掺入多种天然存在和修饰的核苷酸，包括例如终止子核苷酸。

公开(公告)号：US12152256B2

公开(公告)日：2024-11-26

申请号：US17932844

申请日：2022-09-16

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Peter Vander Horn ,  Cheng-Yao Chen ,  Guobin Luo ,  Michael Previte ,  Jamshid Temirov ,  Theo Nikiforov ,  Zhaohui Zhou ,  Hongye Sun ,  Yufang Wang ,  Stefanie Yukiko Nishimura ,  Hongyi Wang ,  Marian Peris ,  Barnett Rosenblum ,  Michael Phelan

IPC: C12Q1/68 ,  C12N9/12 ,  C12Q1/6804 ,  C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6869 ,  C12Q1/6872

Abstract: Provided herein are compositions and systems for use in polymerase-dependent, nucleotide transient-binding methods. The methods are useful for deducing the sequence of a template nucleic acid molecule and single nucleotide polymorphism (SNP) analyses. The methods rely on the fact that the polymerase transient-binding time for a complementary nucleotide is longer compared to that of a non-complementary nucleotide. The labeled nucleotides transiently-binds the polymerase in a template-dependent manner, but does not incorporate. The methods are conducted under any reaction condition that permits transient binding of a complementary or non-complementary nucleotide to a polymerase, and inhibits nucleotide incorporation.

公开(公告)号：US10093969B2

公开(公告)日：2018-10-09

申请号：US15695244

申请日：2017-09-05

Applicant: Life Technologies Corporation

Inventor： Stephen Hendricks ,  David King ,  Lei Xi ,  Marian Peris

IPC: C12Q1/68 ,  C12Q1/6853 ,  C12P19/34 ,  C12Q1/6874 ,  C12Q1/6869 ,  C12Q1/6834 ,  C12Q1/6844 ,  C12Q1/46

Abstract: In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of at least one agent that generates a ligatable terminal 5′ phosphate group by removing an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, an aprataxin enzyme can catalyze removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of an aprataxin enzyme under conditions suitable for ligating nucleic acid ends.

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：US09315861B2

公开(公告)日：2016-04-19

申请号：US14360892

申请日：2012-11-28

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Stephen Hendricks ,  David King ,  Lei Xi ,  Marian Peris

IPC: C12Q1/68 ,  C12P19/34

CPC classification number: C12Q1/46 ,  C12P19/34 ,  C12Q1/6834 ,  C12Q1/6846 ,  C12Q1/6853 ,  C12Q1/6869 ,  C12Q1/6874 ,  C12Q2531/119 ,  C12Q2565/537

Abstract: In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of at least one agent that generates a ligatable terminal 5′ phosphate group by removing an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, an aprataxin enzyme can catalyze removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of an aprataxin enzyme under conditions suitable for ligating nucleic acid ends.

Abstract translation: 在一些实施方案中，用于连接核酸末端的方法包括：在存在可产生可连接的末端5'磷酸基团的至少一种试剂的存在下进行核酸连接反应，通过从核酸的末端5'磷酸脱除腺苷酸基 。 在一些实施方案中，阿伐他汀酶可以催化从核酸的末端5'磷酸脱除腺苷酸基团。 在一些实施方案中，用于连接核酸末端的方法包括：在适于连接核酸末端的条件下，在阿伐他汀酶存在下进行核酸连接反应。