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：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：US20200072826A1

公开(公告)日：2020-03-05

申请号：US16543389

申请日：2019-08-16

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Hua YU ,  Xiaoling YANG ,  Rui ZHENG ,  Chiu Tai Andrew WONG ,  Jeremy GRAY

IPC: G01N33/543 ,  C12Q1/6844

Abstract: The disclosure generally relates to systems, methods, and apparatuses for magnetic bead loading. An example embodiment of the disclosure relates to mixing magnetic beads with sequencing beads to form a solution. The solution containing both beads is injected onto a microchip having a plurality of microwells. The magnetic beads may have larger diameter than the microwell while the sequencing beads may have a smaller diameter, allowing them to enter and reside in the microwell. One or more magnets positioned under the microchip move back and forth across the microchip surface. The magnetic beads form a line and follow the movement of the magnets. During rounds of sweeping, the sequencing beads load into the respective wells. The magnets may be disengaged and the magnetic beads may be washed away after the sequencing beads are loaded.

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：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.