公开(公告)号：US20180057875A1

公开(公告)日：2018-03-01

申请号：US15670885

申请日：2017-08-07

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti ,  Michael P. Weiner ,  Adam R. Abate ,  Tony Hung

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6874 ,  C12Q2537/143 ,  C12Q2563/149

摘要： The present invention relates to systems and methods for sequencing nucleic acids, including sequencing nucleic acids in fluidic droplets. In one set of embodiments, the method employs sequencing by hybridization using droplets such as microfluidic droplets. In some embodiments, droplets are formed which include a target nucleic acid, a nucleic acid probe, and at least one identification element, such as a fluorescent particle. The nucleic acid probes that hybridize to the target nucleic acid are determined, in some instances, by determining the at least one identification element. The nucleic acid probes that hybridize to the target nucleic acid may be used to determine the sequence of the target nucleic acid. In certain instances, the microfluidic droplets are provided with reagents that modify the nucleic acid probe. In some cases, a droplet, such as those described above, is deformed such that the components of the droplets individually pass a target area.

公开(公告)号：US20180023109A1

公开(公告)日：2018-01-25

申请号：US15670977

申请日：2017-08-07

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti ,  Liang-Yin Chu ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George Church

IPC分类号： C12P19/34 ,  B01J13/00 ,  B01F3/08 ,  B01F13/00 ,  C12Q1/68

摘要： The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

公开(公告)号：US20150337371A1

公开(公告)日：2015-11-26

申请号：US14812954

申请日：2015-07-29

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6876 ,  B01J13/04 ,  B01J19/0046 ,  B01J2219/00599 ,  B01J2219/00709 ,  B01J2219/00711 ,  B01J2219/00716 ,  B01J2219/00722 ,  C12Q1/6874

摘要： The present invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

公开(公告)号：US20150336070A1

公开(公告)日：2015-11-26

申请号：US14812946

申请日：2015-07-29

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti

IPC分类号： B01J19/00 ,  C12Q1/68

CPC分类号： C12Q1/6876 ,  B01J13/04 ,  B01J19/0046 ,  B01J2219/00599 ,  B01J2219/00709 ,  B01J2219/00711 ,  B01J2219/00716 ,  B01J2219/00722 ,  C12Q1/6874

摘要： The present invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

公开(公告)号：US20140199730A1

公开(公告)日：2014-07-17

申请号：US14172266

申请日：2014-02-04

申请人： President and Fellows of Harvard College

发明人： Jeremy Agresti ,  Liang-Yin Chu ,  David A. Weitz ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George Church

IPC分类号： C12P19/34

CPC分类号： C12P19/34 ,  B01F3/0807 ,  B01F3/0811 ,  B01F13/0062 ,  B01F13/0071 ,  B01F2215/0037 ,  B01J13/0052 ,  B01J13/0065 ,  B01L3/502784 ,  C12Q1/6834 ,  C12Q1/6848 ,  C12Q1/686 ,  G01N15/1404 ,  G01N2015/1006

摘要： The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

摘要翻译： 本发明通常涉及液滴和/或乳液，例如多重乳液。 在一些情况下，液滴和/或乳液可用于测定中，并且在某些实施方案中，液滴或乳液可被硬化以形成凝胶。 在一些方面，可以使用凝胶进行异质测定。 例如，液滴可以被硬化以形成凝胶，其中液滴包含细胞，DNA或其它合适的物质。 凝胶可以暴露于反应物，并且反应物可以以某种方式与凝胶和/或与细胞，DNA等相互作用。 例如，反应物可以扩散通过凝胶，或者硬化的颗粒可以液化以形成液体状态，允许反应物与细胞相互作用。 作为具体实例，包含在凝胶颗粒内的DNA可以进行PCR（聚合酶链式反应）扩增，例如通过使用能够在形成凝胶时结合凝胶的PCR引物。 当使用PCR扩增DNA时，一些DNA将通过PCR引物与凝胶结合。 PCR反应后，未结合的DNA可以从凝胶中去除，例如通过扩散或洗涤。 因此，可以在本发明的一个实施方案中形成具有结合DNA的凝胶颗粒。

公开(公告)号：US20220396835A1

公开(公告)日：2022-12-15

申请号：US17848909

申请日：2022-06-24

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti

IPC分类号： C12Q1/6876 ,  B01J19/00 ,  C12Q1/6874 ,  B01J13/04

摘要： The present invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

公开(公告)号：US20210348203A1

公开(公告)日：2021-11-11

申请号：US17148796

申请日：2021-01-14

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti ,  Liang-Yin Chu ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George M. Church

IPC分类号： C12P19/34 ,  C12Q1/686 ,  C12Q1/6848 ,  C12Q1/6834 ,  B01F13/00 ,  B01J13/00 ,  G01N15/14 ,  B01F3/08

摘要： The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

公开(公告)号：US10828641B2

公开(公告)日：2020-11-10

申请号：US15496750

申请日：2017-04-25

申请人： President and Fellows of Harvard College

发明人： Christian Boehm ,  Amy Rowat ,  Sarah Koester ,  Jeremy Agresti ,  David A. Weitz

IPC分类号： B01L3/00 ,  B01L7/00 ,  G01N33/50 ,  G01N21/64

摘要： The present invention relates to systems and methods for the arrangement of droplets in pre-determined locations. Many applications require the collection of time-resolved data. Examples include the screening of cells based on their growth characteristics or the observation of enzymatic reactions. The present invention provides a tool and related techniques which addresses this need, and which can be used in many other situations. The invention provides, in one aspect, a tool that allows for stable storage and indexing of individual droplets. The invention can interface not only with microfluidic/microscale equipment, but with macroscopic equipment to allow for the easy injection of liquids and extraction of sample droplets, etc.

公开(公告)号：US20200157593A1

公开(公告)日：2020-05-21

申请号：US16779501

申请日：2020-01-31

申请人： President and Fellows of Harvard College

发明人： David A. Weitz ,  Jeremy Agresti ,  Liang-Yin Chu ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George M. Church

IPC分类号： C12P19/34 ,  C12Q1/686 ,  C12Q1/6848 ,  C12Q1/6834 ,  B01F13/00 ,  B01J13/00 ,  G01N15/14 ,  B01F3/08

摘要： The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

公开(公告)号：US09850526B2

公开(公告)日：2017-12-26

申请号：US14721558

申请日：2015-05-26

申请人： President and Fellows of Harvard College

发明人： Jeremy Agresti ,  Liang-Yin Chu ,  David A. Weitz ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George Church

IPC分类号： C12Q1/68 ,  B01J13/00 ,  C12P19/34 ,  B01F3/08 ,  B01F13/00

CPC分类号： C12P19/34 ,  B01F3/0807 ,  B01F3/0811 ,  B01F13/0062 ,  B01F13/0071 ,  B01F2215/0037 ,  B01J13/0052 ,  B01J13/0065 ,  B01L3/502784 ,  C12Q1/6834 ,  C12Q1/6848 ,  C12Q1/686 ,  G01N15/1404 ,  G01N2015/1006

摘要： The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.