公开(公告)号：US20190062830A1

公开(公告)日：2019-02-28

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

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：US20190062829A1

公开(公告)日：2019-02-28

申请号：US16123140

申请日：2018-09-06

Applicant: Life Technologies Corporation

Inventor： Joseph BEECHEM ,  Theo NIKIFOROV ,  Vi-En CHOONG ,  Xinzhan PENG ,  Guobin LUO ,  Cheng-Yao CHEN ,  Michael PREVITE

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

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：US20170283868A1

公开(公告)日：2017-10-05

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

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.

公开(公告)号：US20170145500A1

公开(公告)日：2017-05-25

申请号：US15423744

申请日：2017-02-03

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Jason MYERS ,  Zhoutao CHEN ,  Devin DRESSMAN ,  Theo NIKIFOROV

IPC: 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.

公开(公告)号：US20160265044A1

公开(公告)日：2016-09-15

申请号：US15166372

申请日：2016-05-27

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: C12Q1/68 ,  C12N9/12 ,  G01N21/64 ,  C12N9/96

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 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.

Abstract translation: 本文公开了包含连接到具有生物活性并可用于多种生物应用的标签的生物分子的缀合物。 例如，本文提供了聚合酶 - 纳米颗粒缀合物，其包括与纳米颗粒连接的聚合酶，其中缀合物具有聚合酶活性。 这样的缀合物可以表现出减少的聚集和改进的沉淀物，其中平均生物分子：纳米颗粒比接近或等于1:1。 本文还公开了制备这种缀合物的改进方法，以及在生物应用中使用这种缀合物的方法和系统，例如核苷酸掺入，引物延伸和单分子测序。

公开(公告)号：US20150353994A1

公开(公告)日：2015-12-10

申请号：US14832772

申请日：2015-08-21

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Jason MYERS ,  Zhoutao CHEN ,  Devin DRESSMAN ,  Theo NIKIFOROV

IPC: 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。

公开(公告)号：US20150094211A1

公开(公告)日：2015-04-02

申请号：US14502201

申请日：2014-09-30

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Peter VANDER HORN ,  Daniel MAZUR ,  Theo NIKIFOROV ,  Mindy LANDES ,  Eileen TOZER

IPC: C12N9/12 ,  C12Q1/68

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.

Abstract translation: 本公开提供了可用于核酸聚合的组合物，方法，试剂盒，系统和装置。 特别地，提供了允许核酸扩增的修饰的聚合酶及其生物活性片段。 在一些方面，本公开提供与参考聚合酶相比具有较低系统误差的修饰聚合酶。 一方面，本公开涉及可用于核酸测序，基因分型，拷贝数变异分析，配对末端测序和其他形式遗传分析的修饰聚合酶。 在一些方面，本公开涉及可用于产生核酸文库或核酸模板的修饰聚合酶。 在一些方面，本公开涉及可以跨越聚合酶的类或家族转移的同源氨基酸突变的鉴定，以提供具有改变的性质的新型聚合酶。