公开(公告)号：US20210346888A1

公开(公告)日：2021-11-11

申请号：US17266911

申请日：2019-08-15

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Iain Clark

IPC: B01L3/00 ,  B01F13/00 ,  B01F3/08

Abstract: The methods described herein provide an improved approach for generating monodispersed droplets. Monodispersed droplets may be effectively obtained by using a plurality of particles to trigger the breakup of a jet, which can include, e.g., flowing in a channel of a microfluidic device a first fluid into a second fluid under stable jetting conditions to provide a jet of the first fluid in the second fluid, wherein the first fluid is immiscible with the second fluid; and introducing a plurality of particles into the jet of the first fluid triggering break-up of the jet of the first fluid and encapsulation of the plurality of particles in a plurality of monodispersed droplets of the first fluid in the second fluid.

公开(公告)号：US11111519B2

公开(公告)日：2021-09-07

申请号：US15015015

申请日：2016-02-03

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  John R. Haliburton ,  Freeman Lan ,  Adam R. Sciambi

IPC: C12Q1/68 ,  C12Q1/6804 ,  C12Q1/6806 ,  C12N15/10 ,  B01L3/00 ,  B01F13/00 ,  B01F5/06 ,  C12Q1/6874 ,  B01L7/00 ,  B01F1/00

Abstract: Microfluidic methods for barcoding nucleic acid target molecules to be analyzed, e.g., via nucleic acid sequencing techniques, are provided. Also provided are microfluidic, droplet-based methods of preparing nucleic acid barcodes for use in various barcoding applications. The methods described herein facilitate high-throughput sequencing of nucleic acid target molecules as well as single cell and single virus genomic, transcriptomic, and/or proteomic analysis/profiling. Systems and devices for practicing the subject methods are also provided.

公开(公告)号：US10745762B2

公开(公告)日：2020-08-18

申请号：US16279905

申请日：2019-02-19

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Dennis Jay Eastburn ,  Adam R. Sciambi

IPC: C12P19/34 ,  C12Q1/6886 ,  C12Q1/6806 ,  B01L3/00 ,  B01L7/00 ,  B01F5/06 ,  B01F13/00 ,  B01F3/08 ,  C12Q1/686 ,  C12Q1/6844

Abstract: The disclosed embodiments relate to method, system and apparatus for synthesizing a target polynucleotide within a droplet. In an exemplary embodiment, the disclosure provides a method of synthesizing a target polynucleotide. The method includes the steps of: contacting a polynucleotide-containing component from a sample with lysis reagents in a droplet, the lysis reagents comprising an enzyme having protease activity, wherein the droplet is encapsulated with an immiscible carrier fluid; moving the droplet into a collection reservoir; incubating the droplet in the collection reservoir for a first duration and then inactivating the enzyme having protease activity; adding to the droplet a nucleic acid synthesis reagent to form a nucleic acid synthesis droplet in the immiscible carrier fluid to form a nucleic acid droplet; and synthesizing the target polynucleotide within the nucleic acid synthesis droplet.

公开(公告)号：US20190169700A1

公开(公告)日：2019-06-06

申请号：US16279905

申请日：2019-02-19

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Dennis Jay Eastburn ,  Adam R. Sciambi

IPC: C12Q1/6886 ,  B01F5/06 ,  B01L7/00 ,  B01L3/00 ,  B01F13/00 ,  B01F3/08 ,  C12Q1/6806 ,  C12Q1/686 ,  C12Q1/6844

Abstract: Methods for the detection of components from biological samples are provided. In certain aspects, the methods may be used to detect and/or quantify specific components in a biological sample, such as tumor cells (e.g., circulating tumor cells). Systems and devices for practicing the subject methods are also provided.

公开(公告)号：US20180237836A1

公开(公告)日：2018-08-23

申请号：US15753132

申请日：2016-08-16

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Freeman Lan ,  Shaun Lim ,  Angus Sidore

IPC: C12Q1/6827 ,  C12P19/34 ,  C12Q1/6844 ,  C40B30/06 ,  C12N9/12

CPC classification number: C12Q1/6827 ,  C12N9/1252 ,  C12P19/34 ,  C12Q1/6844 ,  C12Q2525/179 ,  C12Q2531/119 ,  C12Q2535/122 ,  C12Q2537/16 ,  C12Q2563/159 ,  C40B30/06

Abstract: Methods for non-specifically amplifying a nucleic acid template molecule are provided. The methods may be used to amplify nucleic acid template molecule(s) for sequencing, e.g., for sequencing the genomes of uncultivable microbes or sequencing to identify copy number variation in cancer cells. Aspects of the disclosed methods may include non-specifically amplifying a nucleic acid template molecule, including encapsulating in a microdroplet a nucleic acid template molecule obtained from a biological sample, introducing multiple displacement amplification (MDA) reagents and a plurality of MDA primers into the microdroplet, and incubating the microdroplet under conditions effective for the production of MDA amplification products, wherein the incubating is effective to produce MDA amplification products from the nucleic acid template molecule.

公开(公告)号：US20180216160A1

公开(公告)日：2018-08-02

申请号：US15940850

申请日：2018-03-29

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  John R. Haliburton ,  Freeman Lan ,  Adam R. Sciambi

IPC: C12Q1/6804 ,  C12Q1/6806 ,  C12Q1/6874 ,  C12N15/10 ,  B01L3/00 ,  C12Q1/68 ,  B01F1/00 ,  B01L7/00

Abstract: Microfluidic methods for barcoding nucleic acid target molecules to be analyzed, e.g., via nucleic acid sequencing techniques, are provided. Also provided are microfluidic, droplet-based methods of preparing nucleic acid barcodes for use in various barcoding applications. The methods described herein facilitate high-throughput sequencing of nucleic acid target molecules as well as single cell and single virus genomic, transcriptomic, and/or proteomic analysis/profiling. Systems and devices for practicing the subject methods are also provided.

公开(公告)号：US20180056288A1

公开(公告)日：2018-03-01

申请号：US15520056

申请日：2015-10-21

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  Adam R. Sciambi ,  Russell Cole ,  Zev Jordan Gartner

IPC: B01L3/02 ,  B01L3/00 ,  G01N15/14

Abstract: Methods for delivering discrete entities including, e.g., cells, media or reagents to substrates are provided. In certain aspects, the methods include manipulating and/or analyzing qualities of the entities or biological components thereof. In some embodiments, the methods may be used to create arrays of microenvironments and/or for two and three-dimensional printing of tissues or structures. Systems and devices for practicing the subject methods are also provided.

公开(公告)号：US20170009274A1

公开(公告)日：2017-01-12

申请号：US15015015

申请日：2016-02-03

Applicant: The Regents of the University of California

Inventor： Adam R. Abate ,  John Halliburton ,  Freeman Lan ,  Adam R. Sciambi

IPC: C12Q1/68

CPC classification number: C12Q1/6804 ,  B01F1/00 ,  B01F5/0655 ,  B01F13/0071 ,  B01L3/502707 ,  B01L3/502715 ,  B01L3/502761 ,  B01L3/502784 ,  B01L7/52 ,  B01L2200/0652 ,  B01L2300/021 ,  B01L2300/0645 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/123 ,  B01L2400/0409 ,  B01L2400/0415 ,  B01L2400/0424 ,  B01L2400/043 ,  C12N15/1096 ,  C12Q1/68 ,  C12Q1/6806 ,  C12Q1/6874 ,  C12Q2535/122 ,  C12Q2563/159 ,  C12Q2563/179

Abstract: Microfluidic methods for barcoding nucleic acid target molecules to be analyzed, e.g., via nucleic acid sequencing techniques, are provided. Also provided are microfluidic, droplet-based methods of preparing nucleic acid barcodes for use in various barcoding applications. The methods described herein facilitate high-throughput sequencing of nucleic acid target molecules as well as single cell and single virus genomic, transcriptomic, and/or proteomic analysis/profiling. Systems and devices for practicing the subject methods are also provided.

Abstract translation: 提供了用于条形码核酸靶分子的微流控方法，例如通过核酸测序技术进行分析。 还提供了用于制备用于各种条形码应用的核酸条形码的微流控，基于液滴的方法。 本文描述的方法有助于核酸靶分子以及单细胞和单病毒基因组，转录组学和/或蛋白质组学分析/分析的高通量测序。 还提供了用于实践主题方法的系统和设备。