公开(公告)号：US20220220534A1

公开(公告)日：2022-07-14

申请号：US17708813

申请日：2022-03-30

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Abraham ROSENBAUM ,  Collyn SEEGER ,  Jeremy GRAY ,  Hua YU

IPC: C12Q1/686 ,  C12Q1/6806 ,  C12Q1/6874 ,  C12Q1/6834 ,  C12Q1/6853 ,  C12Q1/6876

Abstract: The present disclosure provides methods, compositions and kits as well as systems for manipulating nucleic acids, including implementing isothermal amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using a pre-seeded solid support. Provided are rapid and efficient methods for generating template nucleic acid molecules comprising specific nucleotide sequence bound to solid support. Such methods can be used, for example, in manipulating nucleic acids in preparation for analysis methods that utilize monoclonal populations of nucleic acids.

公开(公告)号：US20240102079A1

公开(公告)日：2024-03-28

申请号：US18241617

申请日：2023-09-01

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： David JOUN ,  Chieh-Yuan LI ,  Brian REED ,  Craig OBERGFELL ,  Devin DRESSMAN ,  Abraham ROSENBAUM ,  Scott BENSON ,  Andi BROKA ,  Srinka GHOSH

IPC: C12Q1/6806 ,  C12Q1/6844

CPC classification number: C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6846

Abstract: In some embodiments, the disclosure relates generally to compositions, comprising a single reaction mixture containing a plurality of different populations of discrete supports, and a plurality of different populations of target nucleic acids. The single reaction mixture can contain a first population of beads; a second population of beads; a first population of target nucleic acids, where at least two different target nucleic acids in the first population of target nucleic acids can bind to a bead in the first population of beads; and a second population of target nucleic acids, where at least two different target nucleic acids in the second population of target nucleic acids can bind to a bead in the second population of beads. The single reaction mixture can be employed to monoclonally amplify the first target nucleic acids on the first beads, and monoclonally amplify the second target nucleic acids on the second beads.

公开(公告)号：US20190194719A1

公开(公告)日：2019-06-27

申请号：US16183722

申请日：2018-11-07

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Abraham ROSENBAUM ,  Collyn SEEGER ,  Jeremy GRAY ,  Hua YU

IPC: C12Q1/686 ,  C12Q1/6806 ,  C12Q1/6834 ,  C12Q1/6853 ,  C12Q1/6876

CPC classification number: C12Q1/686 ,  C12Q1/6806 ,  C12Q1/6834 ,  C12Q1/6853 ,  C12Q1/6874 ,  C12Q1/6876 ,  C12Q2600/16 ,  C12Q2525/197 ,  C12Q2535/122 ,  C12Q2563/143 ,  C12Q2563/149 ,  C12Q2565/519

Abstract: The present disclosure provides methods, compositions and kits as well as systems for manipulating nucleic acids, including implementing isothermal amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using a pre-seeded solid support. Provided are rapid and efficient methods for generating template nucleic acid molecules comprising specific nucleotide sequence bound to solid support. Such methods can be used, for example, in manipulating nucleic acids in preparation for analysis methods that utilize monoclonal populations of nucleic acids.

公开(公告)号：US20190284597A1

公开(公告)日：2019-09-19

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

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.

公开(公告)号：US20150361481A1

公开(公告)日：2015-12-17

申请号：US14736509

申请日：2015-06-11

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： David JOUN ,  Chieh-Yuan LI ,  Brian REED ,  Craig OBERGFELL ,  Devin DRESSMAN ,  Abraham ROSENBAUM ,  Scott BENSON ,  Andi BROKA ,  Srinka GHOSH

IPC: C12Q1/68

CPC classification number: C12Q1/6806 ,  C12Q1/6844 ,  C12Q2537/143 ,  C12Q2563/149 ,  C12Q2565/537 ,  C12Q2525/191

Abstract: In some embodiments, the disclosure relates generally to compositions, comprising a single reaction mixture containing a plurality of different populations of discrete supports, and a plurality of different populations of target nucleic acids. The single reaction mixture can contain a first population of beads; a second population of beads; a first population of target nucleic acids, where at least two different target nucleic acids in the first population of target nucleic acids can bind to a bead in the first population of beads; and a second population of target nucleic acids, where at least two different target nucleic acids in the second population of target nucleic acids can bind to a bead in the second population of beads. The single reaction mixture can be employed to monoclonally amplify the first target nucleic acids on the first beads, and monoclonally amplify the second target nucleic acids on the second beads.

Abstract translation: 在一些实施方案中，本公开一般涉及组合物，其包含含有多个不同群体的离散载体的单个反应混合物和多个不同的靶核酸群体。 单一反应混合物可以含有第一批珠粒; 第二颗珠子; 靶核酸的第一群体，其中靶核酸的第一群体中的至少两种不同靶核酸可以结合珠粒的第一群体中的珠粒; 和第二靶核酸群，其中靶核酸的第二群体中的至少两种不同靶核酸可以结合到第二群珠中的珠粒。 单个反应混合物可以用于单层扩增第一珠上的第一靶核酸，并单核扩增第二珠上的第二靶核酸。

公开(公告)号：US20220162660A1

公开(公告)日：2022-05-26

申请号：US17667334

申请日：2022-02-08

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.

公开(公告)号：US20190338343A1

公开(公告)日：2019-11-07

申请号：US16514814

申请日：2019-07-17

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： David JOUN ,  Chieh-Yuan LI ,  Brian REED ,  Craig OBERGFELL ,  Devin DRESSMAN ,  Abraham ROSENBAUM ,  Scott BENSON ,  Andi BROKA ,  Srinka GHOSH

IPC: C12Q1/6806 ,  C12Q1/6844

Abstract: In some embodiments, the disclosure relates generally to compositions, comprising a single reaction mixture containing a plurality of different populations of discrete supports, and a plurality of different populations of target nucleic acids. The single reaction mixture can contain a first population of beads; a second population of beads; a first population of target nucleic acids, where at least two different target nucleic acids in the first population of target nucleic acids can bind to a bead in the first population of beads; and a second population of target nucleic acids, where at least two different target nucleic acids in the second population of target nucleic acids can bind to a bead in the second population of beads. The single reaction mixture can be employed to monoclonally amplify the first target nucleic acids on the first beads, and monoclonally amplify the second target nucleic acids on the second beads.