公开(公告)号：US10228348B2

公开(公告)日：2019-03-12

申请号：US15004248

申请日：2016-01-22

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Jene A Golovchenko

IPC: G01N27/447 ,  C12Q1/6869 ,  G01N33/487 ,  B82Y15/00

Abstract: Provided is a solid state support structure including an aperture having a molecular entrance and a molecular exit. A first reservoir is in fluidic communication with the molecular entrance of the aperture and contains a molecule-bearing liquid solution. A second reservoir is in fluidic communication with the molecular exit of the aperture for containing a molecule-bearing liquid solution. A first liquid channel is connected to the first reservoir within less than about 300 microns of the aperture in the support structure and includes molecule-bearing liquid solution for delivery to the first reservoir. A second liquid channel is connected to the second reservoir for accepting molecule-bearing liquid solution from the second reservoir. An electrical connection between the first reservoir and the second reservoir imposes an electrical bias between the first reservoir and the second reservoir for driving the molecule-bearing liquid solution through the aperture in the solid state support structure.

公开(公告)号：US20130313112A1

公开(公告)日：2013-11-28

申请号：US13962141

申请日：2013-08-08

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. DENISON ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

公开(公告)号：US20130270115A1

公开(公告)日：2013-10-17

申请号：US13852606

申请日：2013-03-28

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. DENISON ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

Abstract translation: 本发明涉及通过提供含有足以允许单链单体通过单链多核酸而不是双链多核酸的尺寸的通道的表面来表征目标多核酸的方法; 在表面提供杂交靶标多核酸的来源; 诱导目标多核酸通过通道，由此目标多核酸在其通过之前经历碱基对分离（熔融）; 并且随着靶多核酸相对于通道产生适合于确定目标多核苷酸的单体依赖特性的数据，随时间进行一个或多个测量。

公开(公告)号：US20210278392A1

公开(公告)日：2021-09-09

申请号：US17319254

申请日：2021-05-13

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  C12Q1/6869 ,  G01N27/40

Abstract: A nanopore system provided herein includes first and second fluidic reservoirs in fluidic communication with a nanopore forming a fluidic path between the reservoirs. An enzyme clamp, provided in the first fluidic reservoir, abuts the nanopore and is reversibly bound to a sequential plurality of polymer subunits of a target polymer molecule in ionic solution. The clamp has an outer clamp diameter that is greater than the nanopore diameter. An electrical circuit includes an electrode in each of the reservoirs for applying a voltage bias across the nanopore. A pulse generator is connected in the electrical circuit to apply control pulses across the nanopore to step the clamp along sequential polymer subunits of the target polymer molecule. The system includes no fuel or source of fuel for the clamp. A controller is connected in the electrical circuit for controlling the collection of electrical indications of polymer subunits.

公开(公告)号：US09046483B2

公开(公告)日：2015-06-02

申请号：US13962141

申请日：2013-08-08

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. Denison ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447 ,  C12Q1/68 ,  G01N33/487

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

Abstract translation: 本发明涉及通过提供含有足以允许单链单体通过单链多核酸而不是双链多核酸的尺寸的通道的表面来表征目标多核酸的方法; 在表面提供杂交靶标多核酸的来源; 诱导目标多核酸通过通道，由此目标多核酸在其通过之前经历碱基对分离（熔融）; 并且随着靶多核酸相对于通道产生适合于确定目标多核苷酸的单体依赖特性的数据，随时间进行一个或多个测量。

公开(公告)号：US08986528B2

公开(公告)日：2015-03-24

申请号：US13852606

申请日：2013-03-28

Applicant: President and Fellows of Harvard College

Inventor： Timothy J. Denison ,  Alexis Sauer-Budge ,  Jene A. Golovchenko ,  Amit Meller ,  Eric Brandin ,  Daniel Branton

IPC: G01N27/447 ,  C12Q1/68 ,  G01N33/487

CPC classification number: C12Q1/6869 ,  G01N27/447 ,  G01N33/48721 ,  C12Q2565/631 ,  C12Q2565/607

Abstract: The invention relates to a method for characterizing a target polynucleic acid by providing a surface containing a channel of a dimension sufficient to allow sequential monomer-by-monomer passage of a single-stranded polynucleic acid, but not of a double-stranded polynucleic acid; providing a source of hybridized target polynucleic acid at the surface; inducing passage of the target polynucleic acid through the channel, whereby the target polynucleic acid undergoes base pair separation (melts) prior to its passage; and making one or more measurements over time as the target polynucleic acid moves relative to the channel yielding data suitable to determine a monomer-dependent characteristic of the target polynucleic acid.

公开(公告)号：US12140587B2

公开(公告)日：2024-11-12

申请号：US17319254

申请日：2021-05-13

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  C12Q1/6869 ,  G01N27/40 ,  B82Y35/00

Abstract: A nanopore system provided herein includes first and second fluidic reservoirs in fluidic communication with a nanopore forming a fluidic path between the reservoirs. An enzyme clamp, provided in the first fluidic reservoir, abuts the nanopore and is reversibly bound to a sequential plurality of polymer subunits of a target polymer molecule in ionic solution. The clamp has an outer clamp diameter that is greater than the nanopore diameter. An electrical circuit includes an electrode in each of the reservoirs for applying a voltage bias across the nanopore. A pulse generator is connected in the electrical circuit to apply control pulses across the nanopore to step the clamp along sequential polymer subunits of the target polymer molecule. The system includes no fuel or source of fuel for the clamp. A controller is connected in the electrical circuit for controlling the collection of electrical indications of polymer subunits.

公开(公告)号：US20160139079A1

公开(公告)日：2016-05-19

申请号：US15004248

申请日：2016-01-22

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Jene A. Golovchenko

IPC: G01N27/447

CPC classification number: G01N27/44791 ,  B82Y15/00 ,  C12Q1/6869 ,  G01N27/44743 ,  G01N33/48721 ,  Y10S977/762 ,  C12Q2565/631

Abstract: Provided is a solid state support structure including an aperture having a molecular entrance and a molecular exit. A first reservoir is in fluidic communication with the molecular entrance of the aperture and contains a molecule-bearing liquid solution. A second reservoir is in fluidic communication with the molecular exit of the aperture for containing a molecule-bearing liquid solution. A first liquid channel is connected to the first reservoir within less than about 300 microns of the aperture in the support structure and includes molecule-bearing liquid solution for delivery to the first reservoir. A second liquid channel is connected to the second reservoir for accepting molecule-bearing liquid solution from the second reservoir. An electrical connection between the first reservoir and the second reservoir imposes an electrical bias between the first reservoir and the second reservoir for driving the molecule-bearing liquid solution through the aperture in the solid state support structure.

Abstract translation: 提供了一种固体支持结构，其包括具有分子入口和分子出口的孔。 第一储存器与孔的分子入口流体连通并且包含分子承载液体溶液。 第二储存器与用于容纳分子的液体溶液的孔的分子出口流体连通。 第一液体通道在支撑结构中的孔的小于约300微米内连接到第一储存器，并且包括用于输送到第一储存器的分子承载液体溶液。 第二液体通道连接到第二储存器，用于接收来自第二储存器的分子承载液体溶液。 第一储存器和第二储存器之间的电连接在第一储存器和第二储存器之间施加电偏压，用于驱动分子承载液体溶液穿过固态支撑结构中的孔。

公开(公告)号：US20250060354A1

公开(公告)日：2025-02-20

申请号：US18912894

申请日：2024-10-11

Applicant: President and Fellows Of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  B82Y35/00 ,  C12Q1/6869 ,  G01N27/40

Abstract: A method is provided for deterministically translocating through a nanopore a target polymer molecule of a nucleic acid polymer molecule or a protein polymer molecule. In the method, an enzyme clamp is reversibly bound to a plurality of sequential polymer subunits of the target polymer molecule. The target polymer molecule and the enzyme clamp are disposed at the nanopore. In the method, there is applied a pulse of force operative to deterministically advance the enzyme clamp along the target polymer molecule by no more than one polymer subunit. The pulse of force is then repeatedly applied to cause deterministic translocation of a sequential plurality of polymer subunits of the target polymer molecule through the nanopore.

公开(公告)号：US11035847B2

公开(公告)日：2021-06-15

申请号：US16022594

申请日：2018-06-28

Applicant: President and Fellows of Harvard College

Inventor： Daniel Branton ,  Stephen Jordan Fleming ,  Jene A. Golovchenko

IPC: G01N33/487 ,  B01D71/02 ,  B01D71/74 ,  C12Q1/6869 ,  G01N27/40 ,  B82Y35/00

Abstract: In a method p for controlling translocation of a target polymer molecule through a nanopore, a clamp is reversibly bound to a sequential plurality of polymer subunits along the target polymer molecule length and the molecule and clamp are disposed in an ionic solution that is in fluidic communication with the nanopore. A constant translocation force is applied across the nanopore to induce travel of the target polymer molecule into the nanopore, until the clamp abuts the nanopore aperture and stops further travel of the target polymer molecule into the nanopore. Then a voltage control pulse is applied across the nanopore and/or a thermal control pulse is applied at the nanopore, with a pulse duration that steps the clamp along the target polymer molecule by no more than one polymer subunit in a direction opposite that of travel into the nanopore. No fuel is provided to the clamp.