WO2014028537A1 - Microcapsule compositions and methods - Google Patents

Microcapsule compositions and methods Download PDF

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Publication number
WO2014028537A1
WO2014028537A1 PCT/US2013/054797 US2013054797W WO2014028537A1 WO 2014028537 A1 WO2014028537 A1 WO 2014028537A1 US 2013054797 W US2013054797 W US 2013054797W WO 2014028537 A1 WO2014028537 A1 WO 2014028537A1
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WO
WIPO (PCT)
Prior art keywords
microcapsule
dna
composition
nucleic acid
microcapsules
Prior art date
Application number
PCT/US2013/054797
Other languages
English (en)
French (fr)
Inventor
Benjamin Hindson
Serge Saxonov
Original Assignee
10X Technologies, Inc.
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Filing date
Publication date
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Application filed by 10X Technologies, Inc. filed Critical 10X Technologies, Inc.
Priority to EP13829413.7A priority Critical patent/EP2885418A4/en
Priority to BR112015003354A priority patent/BR112015003354A8/pt
Priority to EP24176310.1A priority patent/EP4397767A3/en
Priority to CN202210540740.9A priority patent/CN114891871A/zh
Priority to CA2881685A priority patent/CA2881685C/en
Priority to CN201380053555.6A priority patent/CN104769127A/zh
Priority to EP21159422.1A priority patent/EP3901273A1/en
Priority to JP2015527549A priority patent/JP6367196B2/ja
Priority to KR1020157006549A priority patent/KR102090851B1/ko
Priority to MX2015001939A priority patent/MX364957B/es
Priority to IN1126DEN2015 priority patent/IN2015DN01126A/en
Priority to AU2013302756A priority patent/AU2013302756C1/en
Priority to CN202010761140.6A priority patent/CN111748607B/zh
Publication of WO2014028537A1 publication Critical patent/WO2014028537A1/en
Priority to US14/316,447 priority patent/US10221442B2/en
Priority to KR1020227005065A priority patent/KR102436171B1/ko
Priority to US14/316,383 priority patent/US20140378345A1/en
Priority to PCT/US2014/044398 priority patent/WO2014210353A2/en
Priority to KR1020227028867A priority patent/KR102642680B1/ko
Priority to EP24158974.6A priority patent/EP4357493A3/en
Priority to BR112015032512A priority patent/BR112015032512A8/pt
Priority to AU2014302277A priority patent/AU2014302277A1/en
Priority to US14/316,398 priority patent/US20150005199A1/en
Priority to MX2015016968A priority patent/MX361481B/es
Priority to KR1020167002243A priority patent/KR102212234B1/ko
Priority to CA2915499A priority patent/CA2915499A1/en
Priority to CN201480047858.1A priority patent/CN105492607B/zh
Priority to CN202311514025.9A priority patent/CN117568449A/zh
Priority to CN201910621839.XA priority patent/CN110592182B/zh
Priority to KR1020217002986A priority patent/KR102366116B1/ko
Priority to US14/316,463 priority patent/US20140378322A1/en
Priority to US14/316,431 priority patent/US20150005200A1/en
Priority to US14/316,416 priority patent/US20140378349A1/en
Priority to EP14817610.0A priority patent/EP3013957B2/en
Priority to JP2016524208A priority patent/JP6563912B2/ja
Priority to EP18189200.1A priority patent/EP3467160A1/en
Priority to IL237156A priority patent/IL237156B/en
Priority to US15/718,764 priority patent/US20180094298A1/en
Priority to US15/718,893 priority patent/US20180051321A1/en
Priority to US15/887,711 priority patent/US20180179580A1/en
Priority to AU2018203012A priority patent/AU2018203012B2/en
Priority to US16/052,486 priority patent/US10323279B2/en
Priority to US16/056,231 priority patent/US11591637B2/en
Priority to US16/231,185 priority patent/US10400280B2/en
Priority to US16/242,962 priority patent/US20190203262A1/en
Priority to US16/246,322 priority patent/US10597718B2/en
Priority to US16/294,769 priority patent/US10450607B2/en
Priority to US16/395,090 priority patent/US10669583B2/en
Priority to US16/435,362 priority patent/US10626458B2/en
Priority to US16/435,417 priority patent/US10752950B2/en
Priority to US16/736,323 priority patent/US12098423B2/en
Priority to US16/844,141 priority patent/US11441179B2/en
Priority to AU2020203602A priority patent/AU2020203602C1/en
Priority to US16/998,425 priority patent/US11035002B2/en
Priority to US16/998,414 priority patent/US11021749B2/en
Priority to AU2020244615A priority patent/AU2020244615B2/en
Priority to US17/314,526 priority patent/US11359239B2/en
Priority to US17/392,610 priority patent/US20220098659A1/en
Priority to AU2022200718A priority patent/AU2022200718B2/en
Priority to US18/186,088 priority patent/US20240002929A1/en
Priority to AU2023203879A priority patent/AU2023203879A1/en
Priority to US18/384,527 priority patent/US20240376519A1/en
Priority to AU2024203235A priority patent/AU2024203235A1/en
Priority to US18/760,903 priority patent/US20250027149A1/en
Priority to US18/945,323 priority patent/US20250066851A1/en
Priority to US18/960,947 priority patent/US20250146047A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/52—Containers specially adapted for storing or dispensing a reagent
    • B01L3/523—Containers specially adapted for storing or dispensing a reagent with means for closing or opening
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1065—Preparation or screening of tagged libraries, e.g. tagged microorganisms by STM-mutagenesis, tagged polynucleotides, gene tags
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2535/00—Reactions characterised by the assay type for determining the identity of a nucleotide base or a sequence of oligonucleotides
    • C12Q2535/122—Massive parallel sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
    • C12Q2563/159—Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components

Definitions

  • the detection and quantification of analytes is important for molecular biology and medical applications such as diagnostics. Genetic testing is particularly useful for a number of diagnostic methods. For example, disorders that are caused by mutations, such as cancer, may be detected or more accurately characterized with DNA sequence information.
  • a starting sample may be a biological sample such as a collection of cells, tissue, or nucleic acids.
  • the sample may need to be lysed or otherwise manipulated in order to permit the extraction of molecules such as DNA.
  • Sample preparation may also involve fragmenting molecules, isolating molecules, and/or attaching unique identifiers to particular fragments of molecules, among other actions.
  • compositions and methods for a microcapsule array device are provided.
  • An aspect of the disclosure provides a composition comprising a first
  • the first microcapsule is degradable upon the application of a stimulus to the first microcapsule; and the first microcapsule comprises an
  • the first microcapsule may comprise a chemical cross-linker.
  • the chemical cross-linker for example, may be a disulfide bond.
  • the composition may comprise a polymer gel, such as, for example a polyacrylamide gel.
  • the first microcapsule may comprise a bead. In some cases, the bead may be a gel bead.
  • the stimulus may be selected from the group consisting of a biological, chemical, thermal, electrical, magnetic, or photo stimulus, and combination thereof.
  • the chemical stimulus may be selected from the group consisting of a change in pH, a change in ion concentration, and a reducing agent.
  • the reducing agent may be, for example, dithiothreitol (DTT) or tm(2-carboxyethyl)
  • TCEP phosphine
  • a second microcapsule may comprise the first microcapsule. Moreover, the second microcapsule may be a droplet.
  • the composition may also comprise a nucleic acid that comprises the oligonucleotide barcode, wherein the nucleic acid comprises a deoxyuridine triphosphate (dUTP).
  • the composition may comprise a polymerase unable to accept a deoxyuridine triphosphate (dUTP).
  • the composition may comprise a target analyte, such as, for example, a nucleic acid.
  • the nucleic acid may be selected from the group consisting of DNA, RNA, dNTPs, ddNTPs, amplicons, synthetic nucleotides, synthetic polynucleotides, polynucleotides,
  • oligonucleotides peptide nucleic acids, cDNA, dsDNA, ssDNA, plasmid DNA, cosmid DNA, High Molecular Weight (MW) DNA, chromosomal DNA, genomic DNA, viral DNA, bacterial DNA, mtDNA (mitochondrial DNA), mRNA, rRNA, tRNA, nRNA, siRNA, snRNA, snoRNA, scaRNA, microRNA, dsRNA, ribozyme, riboswitch and viral RNA.
  • the nucleic acid may be genomic DNA (gDNA).
  • the density of the oligonucleotide barcodes may be at least about 1 ,000,000 oligonucleotide barcodes per the first microcapsule.
  • the oligonucleotide barcode may be coupled to the microcapsule via a chemical cross-linker, such as, for example a disulfide bond.
  • An additional aspect of the disclosure comprises a device comprising a plurality of partitions, wherein: at least one partition of the plurality of partitions comprises a microcapsule comprising an oligonucleotide barcode; and the microcapsule is degradable upon the application of a stimulus to the microcapsule.
  • the partition for example, may be a well or a droplet.
  • the microcapsule comprises a chemical cross-linker such as, for example, a disulfide bond.
  • the microcapsule may comprise a polymer gel such as, for example, a polyacrylamide gel.
  • the microcapsule may comprise a bead. In some cases, the bead may be a gel bead.
  • the stimulus may be selected from the group consisting of a biological, chemical, thermal, electrical, magnetic, or photo stimulus, and a combination thereof.
  • the chemical stimulus may be selected from the group consisting of a change in pH, change in ion concentration, and a reducing agent.
  • the reducing agent for example, may be dithiothreitol (DTT) or tm(2-carboxyethyl) phosphine (TCEP).
  • a nucleic acid may comprise the oligonucleotide barcode and the nucleic acid may comprise a deoxyuridine triphosphate (dUTP).
  • the partition may comprise a polymerase unable to accept a deoxyuridine triphosphate (dUTP).
  • the partition may comprise a target analyte such as, for example, a nucleic acid.
  • the nucleic acid may be selected from the group consisting of DNA, R A, dNTPs, ddNTPs, amplicons, synthetic nucleotides, synthetic polynucleotides,
  • the nucleic acid may be genomic DNA (gDNA).
  • the oligonucleotide barcode may be coupled to the microcapsule via a chemical cross-linker. In some cases, the chemical cross-linker may be a disulfide bond.
  • a further aspect of the disclosure provides a method for sample preparation comprising combining a microcapsule comprising an oligonucleotide barcode and a target analyte into a partition, wherein the microcapsule is degradable upon the application of a stimulus to the microcapsule; and applying the stimulus to the microcapsule to release the oligonucleotide barcode to the target analyte.
  • the partition may be, for example, a well or a droplet.
  • the microcapsule may comprise a polymer gel such as, for example, a polyacrylamide.
  • the microcapsule may comprise a bead.
  • the bead may be a gel bead.
  • the microcapsule may comprise a chemical cross-linker such as, for example, a disulfide bond.
  • the stimulus may be selected from the group consisting of a biological, chemical, thermal, electrical, magnetic, photo stimulus, and a combination thereof.
  • the chemical stimulus may be selected from the group consisting of a change in pH, change in ion concentration, and a reducing agent.
  • the reducing agent may be, for example, dithiothreitol (DTT) or tm(2-carboxyethyl) phosphine (TCEP).
  • DTT dithiothreitol
  • TCEP tm(2-carboxyethyl) phosphine
  • a nucleic acid may comprise the oligonucleotide barcode and the nucleic acid may comprise a deoxyuridine triphosphate (dUTP).
  • the partition may comprise a polymerase unable to accept a deoxyuridine triphosphate (dUTP).
  • the method may also comprise attaching the oligonucleotide barcode to the target analyte. The attaching may be completed, for example, with a nucleic acid amplification reaction.
  • the analyte may be a nucleic acid.
  • the nucleic acid may be selected from the group consisting of DNA, RNA, dNTPs, ddNTPs, amplicons, synthetic nucleotides, synthetic polynucleotides, polynucleotides, oligonucleotides, peptide nucleic acids, cDNA, dsDNA, ssDNA, plasmid DNA, cosmid DNA, High Molecular Weight (MW) DNA, chromosomal DNA, genomic DNA, viral DNA, bacterial DNA, mtDNA (mitochondrial DNA), mRNA, rRNA, tRNA, nRNA, siRNA, snRNA, snoRNA, scaRNA, microRNA, dsRNA, ribozyme, riboswitch and viral RNA.
  • MW Molecular Weight
  • the nucleic acid may be genomic DNA (gDNA).
  • the oligonucleotide barcode may be coupled to the microcapsule via a chemical cross-linker.
  • the chemical cross-linker may be a disulfide bond.
  • a further aspect of the disclosure provides a composition comprising a degradable gel bead, wherein the gel bead comprises at least about 1,000,000 oligonucleotide barcodes. In some cases, the 1,000,000 oligonucleotide barcodes are identical.
  • Fig. 1A is a schematic representation of a microcapsule or inner reagent droplet.
  • Fig. IB is a schematic representation of a microcapsule containing multiple inner reagent droplets.
  • Fig. 2 A is a schematic illustration of a top down view of an exemplary
  • Fig. 2B is a schematic illustration of an exemplary side view of a microcapsule array.
  • FIG. 3 is a schematic illustration of a multi -microcapsule array configuration on a 96-well plate holder.
  • Fig. 4A is a schematic flow diagram representative of a reaction sequence in one microwell of a microwell capsule array.
  • Fig. 4B is similar to 4A, except that it is annotated with examples of methods that can be performed at each step.
  • the present disclosure provides microwell or other partition capsule array devices and methods of using such devices.
  • the device is an assembly of partitions (e.g., microwells, droplets) that are loaded with microcapsules, often at a particular concentration of microcapsules per partition.
  • the devices may be particularly useful to perform sample preparation operations.
  • a device subdivides a sample (e.g., a heterogeneous mixture of nucleic acids, a mixture of cells, etc.) into multiple partitions such that only a portion of the sample is present in each partition.
  • a sample e.g., a heterogeneous mixture of nucleic acids, a mixture of cells, etc.
  • a nucleic acid sample comprising a mixture of nucleic acids may be partitioned such that no more than one strand of (or molecule of) nucleic acid is present in each partition.
  • a cell sample may be partitioned such that no more than one cell is present in each partition.
  • any of a number of different operations may be performed on the subdivided sample within the device.
  • the partitions may include one or more capsules that contain one or more reagents (e.g., enzymes, unique identifiers (e.g., bar codes), antibodies, etc.).
  • the device, a companion device or a user provides a trigger that causes the microcapsules to release one or more of the reagents into the respective partition.
  • the release of the reagent may enable contact of the reagent with the subdivided sample.
  • the reagent is a unique identifier such as a barcode
  • the sample may be tagged with the unique identifier.
  • the tagged sample may then be used in a downstream application such as a sequencing reaction.
  • a variety of different reactions and/operations may occur within a device disclosed herein, including but not limited to: sample partitioning, sample isolation, binding reactions, fragmentation (e.g., prior to partitioning or following partitioning), ligation reactions, and other enzymatic reactions.
  • the device also may be useful for a variety of different molecular biology applications including, but not limited to, nucleic acid sequencing, protein sequencing, nucleic acid quantification, sequencing optimization, detecting gene expression, quantifying gene expression, and single-cell analysis of genomic or expressed markers.
  • the device has numerous medical applications. For example, it may be used for the identification, detection, diagnosis, treatment, staging of, or risk prediction of various genetic and non-genetic diseases and disorders including cancer.
  • FIG. 1A is a schematic of an exemplary microcapsule comprising an internal compartment 120 enveloped by a second layer 130, which is encapsulated by a solid or semi -permeable shell or membrane 110.
  • the shell separates the internal compartment(s) from their immediate environment (e.g., interior of a microwell).
  • the internal compartments, e.g., 120, 130 may comprise materials such as reagents.
  • the reagents 100 may be present in the internal compartment 120. However, in some cases, the reagents are located in the enveloping layer 130 or in both compartments.
  • the microcapsule may release the inner materials, or a portion thereof, following the introduction of a particular trigger. The trigger may cause disruption of the shell layer 110 and/or the internal enveloping layer 130, thereby permitting contact of the internal compartment 100, 120 with the outside environment, such as the cavity of a microwell.
  • the microcapsule may comprise several fluidic phases and may comprise an emulsion (e.g. water-in-oil emulsion, oil-in-water emulsion).
  • a microcapsule may comprise an internal layer 120 that is immiscible with a second layer 130 enveloping the internal layer.
  • the internal layer 120 may comprise an aqueous fluid
  • the enveloping layer 130 may be a non-aqueous fluid such as an oil.
  • the internal layer 120 may comprise a non-aqueous fluid (e.g., oil)
  • the enveloping layer 130 may comprise an aqueous fluid.
  • the microcapsule does not comprise an enveloping second layer.
  • the microcapsule is further encapsulated by a shell layer 110, which may comprise a polymeric material.
  • a microcapsule may comprise a droplet.
  • a microcapsule may be a droplet.
  • the device also may contain a microfluidic element that enables the flow of a sample and/or microcapsules through the device and distribution of the sample and/or microcapsules within the partitions.
  • the microcapsule can comprise multiple compartments.
  • the microcapsule may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 50, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, or 50000 compartments.
  • the microcapsule comprises less than 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 50, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, or 50000 compartments.
  • each compartment, or a subset thereof may also be subdivided into a plurality of additional compartments.
  • each compartment, or subset thereof is subdivided into at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 50, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, or 50000 compartments.
  • each compartment, or subset thereof is further subdivided into less than 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 50, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, or 50000 compartments.
  • each compartment (or some percentage of the total number of
  • compartments may comprise the same reagent or the same combination or reagents. In some cases, each compartment (or some percentage of the total number of compartments) comprises different reagents or a different combination of reagents.
  • the compartments may be configured in a variety of ways. In some cases, the microcapsule may comprise multiple concentric compartments (repeating units of compartments that contain the preceding compartment), often separated by an immiscible layer. In such microcapsules, the reagents may be present in alternating compartments, in every third compartment, or in every fourth compartment.
  • FIG. IB depicts an example of a microcapsule that contains a plurality of smaller microcapsules 140, each containing a reagent.
  • the microcapsule may be encapsulated by an outer shell, often comprising a polymer material 150.
  • the plurality of smaller microcapsules encapsulated within the larger microcapsule may be physically separated by an immiscible fluid 160, thereby preventing mixing of reagents before application of a stimulus and release of reagents into solution.
  • the immiscible fluid is loaded with additional materials or reagents.
  • the plurality of smaller microcapsules are surrounded by a layer of immiscible fluid (e.g., 170) which is further surrounded by a fluid 160 that is miscible with the inner fluid of the microcapsules.
  • the interior microcapsules 180 may comprise an aqueous interior enveloped by an immiscible (e.g., oil) layer, that is further surrounded by an aqueous layer 160.
  • the miscible compartments e.g., 160 and 180
  • the microcapsule may comprise a polymeric shell (see, e.g., Figures 1 and 2) or multiple polymeric shells.
  • the microcapsule may comprise multiple polymeric shells layered on top of each other.
  • individual compartments within a microcapsule comprise a polymeric shell, or a subset of the compartments may comprise a polymeric shell.
  • all or some of the smaller compartments 140 in Figure IB may comprise a polymeric shell that separates them from the fluidic interior 160.
  • the microcapsule may be designed so that a particular reagent is contained within a compartment that has a polymerized shell, while a different reagent is within a compartment that is simply enveloped by an immiscible liquid.
  • a reagent that is desired to be released upon a heat trigger may be contained within the
  • paramagnetic particles may be incorporated into the capsule shell wall.
  • a magnet or electric field may then be used to position the capsule to a desired location.
  • a magnetic field e.g., high frequency alternating magnetic field
  • the incorporated paramagnetic particles may then transform the energy of the magnetic field into heat, thereby triggering rupture of the capsule.
  • microcapsule component of a device of this disclosure may provide for the controlled and/or timed release of reagents for sample preparation of an analyte.
  • Microcapsules may be used in particular for controlled release and transport of varying types of chemicals, ingredients, pharmaceuticals, fragrances etc..., including particularly sensitive reagents such as enzymes and proteins (see, e.g., D. D. Lewis, "Biodegradable Polymers and Drug Delivery Systems", M. Chasin and R. Langer, editors (Marcel Decker, New York, 1990); J. P. McGee et al, J. Control. Release 34 (1995), 77).
  • Microcapsules may also provide a means for delivery of reagents in discrete and definable amounts. Microcapsules may be used to prevent premature mixing of reagents with the sample, by segregating the reagents from the sample. Microcapsules also may ease handling of - and limit contacts with— particularly sensitive reagents such as enzymes, nucleic acids and other chemicals used in sample preparation.
  • Microcapsules of a device of this disclosure may be prepared by numerous methods and processes. Preparative techniques may include pan coating, spray drying, centrifugal extrusion, emulsion-based methods, and/or microfluidic techniques. Typically, a method for preparation is chosen based on the desired characteristics of the
  • microcapsule For example, shell wall thickness, permeability, chemical composition of the shell wall, mechanical integrity of the shell wall and capsule size may be taken into consideration when choosing a method. Methods of preparation may also be selected based on the ability to incorporate specific materials within the capsule such as whether the core materials (e.g., fluids, reagents, etc.) are aqueous, organic or inorganic.
  • core materials e.g., fluids, reagents, etc.
  • preparation methods can affect the shape and size of the microcapsule.
  • a capsule's shape (e.g., spherical, ellipsoidal, etc.) may depend on the shape of the droplet in the precursor liquid which may be determined by the viscosity and surface tension of the core liquid, direction of flow of the emulsion, the choice of surfactants used in droplet stabilization, as well as physical confinement such as preparations made in a microchannel or capillary of a particular size (e.g., a size requiring distortion of the microcapsule in order for the microcapsule to fit within the microchannel or capillary.
  • Microcapsules may be prepared through emulsification polymerization, a process in which monomer units at an aqueous/organic interface in an emulsion polymerize to form a shell. Reagents are mixed with the aqueous phase of the biphasic mixture.
  • Vigorous shaking, or sonication of the mixture creates droplets containing reagents, which are encased by a polymeric shell.
  • microcapsules may be prepared through layer-by-layer assembly, a process in which negatively and positively charged polyelectrolytes are deposited onto particles such as metal oxide cores. Electrostatic interactions between polyelectrolytes create a polymeric shell around the core. The core can be subsequently removed via addition of acid, resulting in a semi-permeable hollow sphere which can be loaded with various reagents.
  • microcapsules may be prepared through coacervation, a process in which two oppositely charged polymers in aqueous solution become entangled to form a neutralized polymer shell wall.
  • One polymer may be contained within an oil phase, while the other, of opposite charge is contained in an aqueous phase.
  • This aqueous phase may contain reagents to be encapsulated.
  • the attraction of one polymer for another can result in the formation of coascervates.
  • gelatin and gum Arabic are components of this preparative method.
  • Microcapsules also may be prepared through internal phase separation, a process in which a polymer is dissolved in a solvent mixture containing volatile and nonvolatile solvents. Droplets of the resultant solution are suspended in an aqueous layer, which is stabilized by continual agitation and the use of surfactants. This phase may contain reagents to be encapsulated. When the volatile solvent evaporates, the polymers coalesce to form a shell wall. In some cases, polymers such as polystyrene, poly(methyl methacrylate) and poly(tetrahydrofuran) are used to form shell walls.
  • Microcapsules also may be prepared through flow focusing methods, a process in which a microcapillary device is used to generate double emulsions containing a single internal droplet encased in a middle fluid which is then dispersed to an outer fluid.
  • the inner droplet may contain reagents to be encapsulated.
  • the middle fluid becomes the shell wall, which can be formed via cross-linking reactions.
  • Microcapsules may comprise a variety of materials with a wide range of chemical characteristics. Generally, the microcapsules comprise materials with the ability to form microcapsules of a desired shape and size and that are compatible with the reagents to be stored in the microcapsules.
  • Microcapsules may comprise a wide range of different polymers including but not limited to: polymers, heat sensitive polymers, photosensitive polymers, magnetic polymers, pH sensitive polymers, salt-sensitive polymers, chemically sensitive polymers, polyelectrolytes, polysaccharides, peptides, proteins, and/or plastics.
  • Polymers may include but are not limited to materials such as poly(N-isopropylacrylamide)
  • PNIPAAm poly(styrene sulfonate)
  • PSS poly(allyl amine)
  • PAAm poly(acrylic acid)
  • PAA poly(ethylene imine)
  • PEI poly(diallyldimethyl-ammonium chloride)
  • PDADMAC poly(pyrolle)
  • PVPON polyvinylpyrrolidone
  • PMAA poly(methacrylic acid)
  • PMMA poly(methyl methacrylate)
  • PS polystyrene
  • PTHF poly(tetrahydrofuran)
  • PHF poly(phthaladehyde)
  • PHF poly(hexyl viologen)
  • PLL poly(L-lysine)
  • PARG poly(lactic- co-glycolic acid)
  • PLGA poly(lactic- co-glycolic acid)
  • a microcapsule may be prepared from heat sensitive polymers and/or may comprise one or more shells comprising such heat-sensitive polymers.
  • the heat-sensitive polymer may be stable under conditions used for storage or loading. Upon exposure to heat, the heat- sensitive polymer components may undergo depolymerization, resulting in disruption to the integrity of the shell and release of the inner materials of the microcapsule (and/or of the inner microcapsules) to the outside environment (e.g., the interior of a microwell).
  • Exemplary heat-sensitive polymers may include, but are not limited to NIPAAm or PNIPAM hydrogel.
  • the microcapsules may also comprise one or more types of oil.
  • Exemplary oils include but are not limited to hydrocarbon oils, fluorinated oils, fluorocarbon oils, silicone oils, mineral oils, vegetable oils, and any other suitable oil.
  • the microcapsules may also comprise a surfactant, such as an emulsifying surfactant.
  • exemplary surfactants include, but are not limited to, cationic surfactants, non- ionic surfactants, anionic surfactants, hydrocarbon surfactants or fluorosurfactants.
  • the surfactant may increase the stability of one or more components of the microcapsule, such as an inner compartment that comprises an oil.
  • the microcapsules may comprise an inner material that is miscible with materials external to the capsule.
  • the inner material may be an aqueous fluid and the sample within the microwell may also be in an aqueous fluid.
  • the microcapsule may comprise powders or nanoparticles that are miscible with an aqueous fluid.
  • the microcapsule may comprise such powders or nanoparticles in an inner compartment. Upon disruption of the microcapsule, such powders or nanoparticles are released into the external environment (e.g., interior of microwell) and may mix with an aqueous fluid (e.g., an aqueous sample fluid).
  • an aqueous fluid e.g., an aqueous sample fluid
  • the microcapsule may comprise a material that is immiscible with the surrounding environment (e.g., interior of microwell, sample fluid).
  • the phase separation between the inner and outer components may promote mixing, such as mixing of the inner components with the surrounding fluid.
  • a pressure or force is also released that promotes mixing of internal and external components.
  • the microcapsules may also comprise a polymer within the interior of the capsule.
  • this polymer may be a porous polymer bead that may entrap reagents or combinations of reagents.
  • this polymer may be a bead that has been previously swollen to create a gel.
  • Examples of polymer based gels that may be used as inner emulsions of capsules may include, but are not limited to sodium alginate gel, or poly acrylamide gel swelled with oligonucleotide bar codes or the like.
  • a microcapsule may be a gel bead comprising any of the polymer based gels described herein.
  • Gel bead microcapsules may be generated, for example, by encapsulating one or more polymeric precursors into droplets. Upon exposure of the polymeric precursors to an accelerator (e.g., tetramethylethylenediamine (TEMED)), a gel bead may be generated.
  • an accelerator e.g., tetramethylethylenediamine (TEMED)
  • Analytes and/or reagents such as oligonucleotide barcodes, for example, may be coupled/immobilized to the interior surface of a gel bead (e.g., the interior accessible via diffusion of an oligonucleotide barcode and/or materials used to generate an oligonucleotide barcode and/or materials used to generate an oligonucleotide barcode and/or materials used to generate an
  • oligonucleotide barcode and/or the outer surface of a gel bead or any other microcapsule described herein.
  • Coupling/immobilization may be via any form of chemical bonding (e.g., covalent bond, ionic bond) or physical phenomena (e.g., Van der Waals forces, dipole-dipole interactions, etc.).
  • coupling/immobilization of a reagent to a gel bead or any other microcapsule described herein may be reversible, such as, for example, via a labile moiety (e.g., via a chemical cross-linker, including chemical cross- linkers described herein).
  • the labile moiety may be cleaved and the immobilized reagent set free.
  • the labile moiety is a disulfide bond.
  • an oligonucleotide barcode is
  • the labile moiety may be included as part of a gel bead or microcapsule, as part of a chemical linker that links a reagent or analyte to a gel bead or microcapsule, and/or as part of a reagent or analyte.
  • a gel bead or any other type of microcapsule described herein may contain varied numbers of reagents.
  • the density of a reagent per microcapsule may vary depending on the particular microcapsule utilized and the particular reagent.
  • a microcapsule or gel bead may comprise at least about 1; 10; 100; 1,000; 10,000; 100,000; 1,000,000; 5,000,000; 10,000,000, 50,000,000; 100,000,000; 500,000,000; or
  • a gel bead may comprise identical oligonucleotide barcodes or may comprise differing oligonucleotide barcodes.
  • the microcapsule may comprise one or more materials that create a net neutral, negative or positive charge on the outer shell wall of the capsule.
  • the charge of a capsule may aid in preventing or promoting aggregation or clustering of particles, or adherence or repulsion to parts of the device.
  • the microcapsule may comprise one or more materials that cause the outer shell wall of the capsule to be hydrophilic or hydrophobic.
  • a hydrophilic material that may be used for capsule shell walls may be poly(N-isopropylacrylamide).
  • a hydrophobic material that may be used for capsule shell walls may be polystyrene.
  • a hydrophilic shell wall may aid in wicking of the capsule into wells comprising aqueous fluid.
  • a microcapsule may be any of a number of sizes or shapes.
  • the shape of the microcapsule may be spherical, ellipsoidal, cylindrical, hexagonal or any other symmetrical or non-symmetrical shape. Any cross-section of the microcapsule may also be of any appropriate shape, include but not limited to: circular, oblong, square, rectangular, hexagonal, or other symmetrical or non-symmetrical shape.
  • the microcapsule may be of a specific shape that complements an opening (e.g., surface of a microwell) of the device.
  • the microcapsule may be spherical and the opening of a microwell of the device may be circular.
  • the microcapsules may be of uniform size (e.g., all of the microcapsules are the same size) or heterogeneous size (e.g., some of the microcapsules are of different sizes).
  • a dimension (e.g., diameter, cross-section, side, etc.) of a microcapsule may be at least about 0.001 ⁇ , .01 ⁇ , 0.1 ⁇ , 0.5 ⁇ , 1 ⁇ , 5 ⁇ , 10 ⁇ , 50 ⁇ , 100 ⁇ , 200 ⁇ , 300 ⁇ , 400 ⁇ , 500 ⁇ , 600 ⁇ , 700 ⁇ , 800 ⁇ , 900 ⁇ or 1 nm.
  • the microcapsule comprises a microwell that is at most about 0.001 ⁇ , .01 ⁇ , 0.1 ⁇ , 0.5 ⁇ , 1 ⁇ , 5 ⁇ , 10 ⁇ , 50 ⁇ , 100 ⁇ , 200 ⁇ , 300 ⁇ , 400 ⁇ , 500 ⁇ , 600 ⁇ , 700 ⁇ , 800 ⁇ , 900 ⁇ or 1 nm.
  • microcapsules are of a size and/or shape so as to allow a limited number of microcapsules to be deposited in individual partitions (e.g., microwells, droplets) of the microcapsule array.
  • Microcapsules may have a specific size and/or shape such that exactly or no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 capsules fit into an individual microwell; in some cases, on average 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 capsules fit into an individual microwell. In still further cases, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 100, 500, or 1000 capsules fit into an individual microwell.
  • the devices provided herein may comprise free reagents and/or reagents encapsulated into microcapsules.
  • the reagents may be a variety of molecules, chemicals, particles, and elements suitable for sample preparation reactions of an analyte.
  • a microcapsule used in a sample preparation reaction for DNA sequencing of a target may comprise one or more of the following reagents: enzymes, restriction enzymes (e.g., multiple cutters), ligase, polymerase (e.g., polymerases that do and do not recognize dUTPs and/or uracil), fluorophores, oligonucleotide barcodes, buffers, deoxynucleotide triphosphates (dNTPs) (e.g.deoxyadenosine triphosphate (dATP), deoxycitidine triphosphate (dCTP), deoxyguanosine triphosphate (dGTP), deoxythymidine triphosphate (dTTP), deoxyuridine triphosphate (dUTP)), deoxynucleotide triphosphates (ddNTPs) and the like.
  • enzymes e.g., restriction enzymes (e.g., multiple cutters), ligase, polymerase (e.g
  • a microcapsule used in a sample preparation reaction for single cell analysis may comprise reagents such as one or more of the following reagents: lysis buffer, detergent, fluorophores, oligonucleotide barcodes, ligase, proteases, heat activatable proteases, protease or nuclease inhibitors, buffer, enzymes, antibodies, nanoparticles, and the like.
  • reagents such as one or more of the following reagents: lysis buffer, detergent, fluorophores, oligonucleotide barcodes, ligase, proteases, heat activatable proteases, protease or nuclease inhibitors, buffer, enzymes, antibodies, nanoparticles, and the like.
  • Exemplary reagents include, but are not limited to: buffers, acidic solution, basic solution, temperature-sensitive enzymes, pH-sensitive enzymes, light-sensitive enzymes, metals, metal ions, magnesium chloride, sodium chloride, manganese, aqueous buffer, mild buffer, ionic buffer, inhibitor, enzyme, protein, nucleic acid, antibodies, saccharides, lipid, oil, salt, ion, detergents, ionic detergents, non-ionic detergents, oligonucleotides, nucleotides, dNTPs, ddNTPs, deoxyribonucleic acid (DNA), ribonucleic acid (R A), peptide nucleic acids, circular DNA (cDNA), double-stranded DNA (dsDNA), single- stranded DNA (ssDNA), plasmid DNA, cosmid DNA, chromosomal DNA, genomic DNA (gDNA), viral DNA, bacterial DNA, mtDNA (mitochondrial DNA), messenger aqueous
  • a microcapsule comprises a set of reagents that have a similar attribute (e.g., a set of enzymes, a set of minerals, a set of oligonucleotides, a mixture of different bar-codes, a mixture of identical bar-codes).
  • a microcapsule comprises a heterogeneous mixture of reagents.
  • the heterogeneous mixture of reagents comprises all components necessary to perform a reaction.
  • such mixture comprises all components necessary to perform a reaction, except for 1, 2, 3, 4, 5, or more components necessary to perform a reaction.
  • Reagents may be pre-loaded into the device (e.g., prior to introduction of analyte) or post-loaded into the device. They may be loaded directly into the device; or, in some cases, the reagents are encapsulated into a microcapsule that is loaded into the device. In some cases, only microcapsules comprising reagents are introduced. In other cases, both free reagents and reagents encapsulated in microcapsules are loaded into the device, either sequentially or concurrently.
  • reagents are introduced to the device either before or after a particular step.
  • a lysis buffer reagent may be introduced to the device following partitioning of a cellular sample into multiple partitions (e.g., microwells, droplets) within the device.
  • reagents and/or microcapsules comprising reagents are introduced sequentially such that different reactions or operations occur at different steps.
  • the reagents (or microcapsules) may be also be loaded at steps interspersed with a reaction or operation step.
  • microcapsules comprising reagents for fragmenting molecules (e.g., nucleic acids) may be loaded into the device, followed by a fragmentation step, which may be followed by loading of microcapsules comprising reagents for ligating bar-codes (or other unique identifiers, e.g., antibodies) and subsequent ligation of the bar-codes to the fragmented molecules. Additional methods of loading reagents are described further herein in other sections.
  • one or more unique molecular identifiers are used as sample preparation reagents. These molecules may comprise a variety of different forms such as oligonucleotide bar codes, antibodies or antibody fragments, fluorophores, nanop articles, and other elements or combinations thereof. Depending upon the specific application, molecular barcodes may reversibly or irreversibly bind to the target analyte and allow for identification and/or quantification of individual analytes after recovery from a device after sample preparation.
  • a device of this disclosure may be applicable to nucleic acid sequencing, protein detection, single molecule analysis and other methods that require a) precise measurement of the presence and amount of a specific analyte b) multiplex reactions in which multiple analytes are pooled for analysis.
  • a device of this disclosure may utilize the microwells of the microwell array or other type of partition (e.g., droplets) to physically partition target analytes. This physical partitioning allows for individual analytes to acquire one or more molecular barcodes. After sample preparation, individual analytes may be pooled or combined and extracted from a device for multiplex analysis.
  • Molecular barcodes may allow for the identification and quantification of individual molecules even after pooling of a plurality of analytes.
  • molecular barcodes may permit the sequencing of individual nucleic acids, even after the pooling of a plurality of different nucleic acids.
  • Oligonucleotide barcodes may be particularly useful in nucleic acid sequencing.
  • an oligonucleotide barcode may comprise a unique sequence (e.g., a barcode sequence) that gives the oligonucleotide barcode its identifying functionality.
  • the unique sequence may be random or non-random. Attachment of the barcode sequence to a nucleic acid of interest may associate the barcode sequence with the nucleic acid of interest.
  • the barcode may then be used to identify the nucleic acid of interest during sequencing, even when other nucleic acids of interest (e.g., comprising different barcodes) are present.
  • an attached barcode may be used to identify fragments as belonging to the nucleic acid of interest during sequencing.
  • An oligonucleotide barcode may consist solely of a unique barcode sequence or may be included as part of an oligonucleotide of longer sequence length.
  • Such an oligonucleotide may be an adaptor required for a particular sequencing chemistry and/or method.
  • such adaptors may include, in addition to an oligonucleotide barcode, immobilization sequence regions necessary to immobilize (e.g., via
  • an adaptor to a solid surface (e.g., solid surfaces in a sequencer flow cell channel); sequence regions required for the binding of sequencing primers; and/or a random sequence (e.g., a random N-mer) that may be useful, for example, in random amplification schemes.
  • An adaptor can be attached to a nucleic acid to be sequenced, for example, by amplification, ligation, or any other method described herein.
  • an oligonucleotide barcode, and/or a larger oligonucleotide comprising an oligonucleotide barcode may comprise natural nucleic acid bases and/or may comprise non-natural bases.
  • the oligonucleotide may comprise the natural DNA bases adenine, guanine, cytosine, and thymine and/or may comprise non-natural bases such as uracil.
  • reagent loaded microcapsules may be loaded into a device using a variety of methods.
  • Microcapsules in some instances, may be loaded as 'dry capsules.'
  • capsules may be separated from a liquid phase using various techniques, including but not limited to differential centrifugation, evaporation of the liquid phase, chromatography, filtration and the like.
  • 'Dry capsules' may be collected as a powder or particulate matter and then deposited into microwells of the microwell array. Loading 'dry capsules' may be a preferred method in instances in which loading of 'wet capsules,' leads to inefficiencies of loading such as empty wells and poor distribution of microcapsules across the microwell array.
  • Reagent-loaded microcapsules may also be loaded into a device when the microcapsules are within a liquid phase, and thereby loaded as 'wet capsules.'
  • microcapsules may be suspended in a volatile oil such that the oil can be removed or evaporated, leaving only the dry capsule in the well.
  • Loading 'wet capsules' may be a preferred method in some instances in which loading of dry capsules leads to inefficiencies of loading, such as microcapsule clustering, aggregation and poor distribution of microcapsules across the microwell array. Additional methods of loading reagents and microcapsules are described in other sections of this disclosure.
  • the microcapsules also may have a particular density.
  • the microcapsules are less dense than an aqueous fluid (e.g., water); in some cases, the microcapsules are denser than an aqueous fluid (e.g., water).
  • the microcapsules are less dense than a non-aqueous fluid (e.g., oil); in some cases, the microcapsules are denser than a non-aqueous fluid (e.g., oil).
  • Microcapsules may comprise a density at least about 0.05 g/cm 3 , 0.1 cm 3 , 0.2 g/ cm 3 , 0.3 g/ cm 3 , 0.4 g/ cm 3 , 0.5 g/ cm 3 , 0.6 g/ cm 3 , 0.7 g/cm 3 , 0.8 g/cm 3 , 0.81 g/cm 3 , 0.82 g/cm 3 , 0.83 g/cm 3 , 0.84 g/cm 3 , 0.85 g/cm 3 , 0.86 g/cm 3 , 0.87 g/cm 3 , 0.88 g/cm 3 , 0.89 g/cm 3 , 0.90 g/cm 3 , 0.91 g/cm 3 , 0.92 g/cm 3 , 0.93 g/cm 3 , 0.94 g/cm 3 , 0.95 g/cm 3 , 0.96 g/cm 3 , 0.
  • microcapsule densities may be at most about 0.7 g/cm 3 , 0.8 g/cm 3 , 0.81 g/cm 3 , 0.82 g/cm 3 , 0.83 g/cm 3 , 0.84 g/cm 3 , 0.85 g/cm 3 , 0.86 g/cm 3 , 0.87 g/cm 3 , 0.88 g/cm 3 , 0.89 g/cm 3 , 0.90 g/cm 3 , 0.91 g/cm 3 , 0.92 g/cm 3 , 0.93 g/cm 3 , 0.94 g/cm 3 , 0.95 g/cm 3 , 0.96 g/cm 3 , 0.97 g/cm 3 , 0.98 g/cm 3 , 0.99 g/cm 3 , 1.00 g/cm 3 , 1.05 g/cm 3 , 1.1 g/cm3 ,
  • a device of this disclosure may be a microwell array comprising a solid plate containing a plurality of holes, cavities or microwells in which microcapsules and/or analytes are deposited.
  • a fluidic sample or analyte
  • the microwells may comprise microcapsules when the sample is introduced; or, in some cases, the microcapsules are introduced into the microwells following introduction of the sample.
  • FIG. 2A depicts a prototype microwell array; a sideview is depicted in Figure 2B.
  • the microwell array may include a plate 220 that can be made of any suitable material commonly used in a chemical laboratory, including fused silica, soda lima glass, borosilicate glass, PMMA, sapphire, silicon, germanium, cyclic olefin copolymer and cyclic polymer, polyethylenes, polypropylenes, polyacrylates, polycarbonates, plastics, Topas, and other suitable substrates known in the art.
  • the plate 220 may initially be a flat solid plate comprising a regular pattern of microwells 270.
  • microwells may be formed by drilling or chemical dissolution or any other suitable method of machining; however, plates with a desired hole pattern are preferably molded, e.g. by injection- molding, embossing, or using a suitable polymer, such as cyclic olefin copolymer.
  • the microwell array may comprise an inlet (200 and 240) and/or an outlet (210 and 260); in some cases, the microwell array comprises multiple inlets and/or outlets.
  • a sample (or analyte) or microcapsules may be introduced to the device via the inlet.
  • Solutions containing analytes, reagents and/or microcapsules may be manually applied to the inlet port 200 and 240 (or to a conduit attached to the inlet port) via a pipette.
  • a liquid handling device is used to introduce analytes, reagents, and/or
  • the inlet port is connected to a reservoir comprising microcapsules or analytes.
  • the inlet port may be attached to a flow channel 250 that permits distribution of the analyte, sample, or microcapsules to the microwells in the device.
  • the inlet port may be used to introduce to the device a fluid (e.g., oil, aqueous) that does not contain microcapsules or analyte, such as a carrier fluid.
  • the carrier fluid may be introduced via the inlet port before, during, or following the introduction of analyte and/or microcapsules.
  • the same sample may be introduced via the multiple inlets, or each inlet may convey a different sample.
  • one inlet may convey a sample or analyte to the microwells, while a different inlet conveys free reagents and/or reagents encapsulated in microcapsules to the device.
  • the device may have at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 inlets and/or outlets.
  • solutions containing microcapsules and/or analytes may be pulled through the device via a vacuum manifold attached to the outlet port 210 and 260.
  • a vacuum manifold may apply a negative pressure to the device.
  • a positive pressure is used to move sample, analytes, and/or microcapsules through the device.
  • the area, length, and width of surfaces of 230 according to this disclosure may be varied according to the requirements of the assay to be performed. Considerations may include, for example, ease of handling, limitations of the material(s) of which the surface is formed, requirements of detection or processing systems, requirements of deposition systems (e.g. microfluidic systems), and the like.
  • the thickness may comprise a thickness of at least about 0.001 mm, 0.005 mm, 0.01 mm, 0.05 mm, 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm, 1.0 mm, 2.0 mm, 3.0 mm, 4.0 mm, 5.0 mm, 6.0 mm, 7.0 mm, 8.0 mm, 9.0 mm, 10.0 mm, 11 mm, 12 mm, 13 mm, 14 mm, or 15 mm.
  • microcapsule thickness may be at most 0.001 mm, 0.005 mm, 0.01 mm, 0.05 mm, 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm, 1.0 mm, 2.0 mm, 3.0 mm, 4.0 mm, 5.0 mm, 6.0 mm, 7.0 mm, 8.0 mm, 9.0 mm, 10.0 mm, 11 mm, 12 mm, 13 mm, 14 mm, or 15 mm.
  • the microwells 270 can be any shape and size suitable for the assay performed.
  • the cross-section of the microwells may have a cross-sectional dimension that is circular, rectangular, square, hexagonal, or other symmetric or non-symmetric shape.
  • the shape of the microwell may be cylindrical, cubic, conical, frustoconical, hexagonal or other symmetric or non-symmetric shape.
  • the diameter of the microwells 270 may be determined by the size of the wells desired and the available surface area of the plate itself.
  • Exemplary microwells comprise diameters of at least 0.01 ⁇ , 0.1 ⁇ , 0.2 ⁇ , 0.3 ⁇ , 0.4 ⁇ , 0.5 ⁇ , 1 ⁇ , 10 ⁇ , 25 ⁇ , 50 ⁇ , 75 ⁇ , 100 ⁇ , 200 ⁇ m, 300 ⁇ , 400 ⁇ m, 500 ⁇ , 600 ⁇ m, 700 ⁇ m, 800 ⁇ , 900 ⁇ m, 1.0 mm.
  • microwell diameters may comprise at most 0.01 ⁇ , 0.1 ⁇ , 0.2 ⁇ , 0.3 ⁇ , 0.4 ⁇ m, 0.5 ⁇ , 1 ⁇ m, 10 ⁇ , 25 ⁇ m, 50 ⁇ m, 75 ⁇ , 100 ⁇ m, 200 ⁇ , 300 ⁇ m, 400 ⁇ m, 500 ⁇ , 600 ⁇ , 700 ⁇ , 800 ⁇ , 900 ⁇ m or 1.0 mm.
  • the capacity (or volume) of each well can be a measure of the height of the well (the thickness of the plate) and the effective diameter of each well.
  • the capacity of an individual well may be selected from a wide range of volumes.
  • the device may comprise a well (or microwell) with a capacity of at least 0.001 fL, .01 fL, 0.1 fL, 0.5 fL, 1 fL, 5 fL, 10 fL, 50 fL, 100 fL, 200 fL, 300 fL, 400 fL, 500 fL, 600 fL, 700 fL, 800 fL, 900 fL, 1 pL, 5 pL, 10 pL, 50 pL, 100 pL, 200 pL, 300 pL, 400 pL, 500 pL, 600 pL, 700 pL, 800 pL, 900 pL, 1 nL, 5 nL, 10 nL, 50 n
  • the microcapsule comprises a microwell that is less than 0.001 fL, .01 fL, 0.1 fL, 0.5 L, 5 fL, 10 fL, 50 fL, 100 fL, 200 fL, 300 fL, 400 fL, 500 fL, 600 fL, 700 fL, 800 fL, 900 fL, 1 pL, 5 pL, 10 pL, 50 pL, 100 pL, 200 pL, 300 pL, 400 pL, 500 pL, 600 pL, 700 pL, 800 pL, 900 pL, 1 nL, 5 nL, 10 nL, 50 nL, 100 nL, 200 nL, 300 nL, 400 nL, 500 nL, 1 uL, 50 uL, or 100 uL.
  • volume of fluid in different microwells in the array may vary by at least (or at most) plus or minus 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, or 1000% across a set of microwells.
  • a microwell may comprise a volume of fluid that is at most 80% of the fluid volume within a second microwell.
  • the microwell array may comprise a range of well densities.
  • a plurality of microwells may have a density of at least about 2,500 wells/cm , at least about 1,000 wells/cm .
  • the plurality of wells may have a density of at least 10 wells/cm 2.
  • the well density may comprise at least 10 wells/cm 2 , 50 wells/cm 2 , 100 wells/cm 2 , 500 wells/cm 2 , 1000 wells/cm 2 , 5000 wells/cm 2 , 10000 wells/cm 2 50000 wells/cm 2 or 100000 wells/cm 2". In other cases, the well density may be less than 100000 wells/cm 2 , 10000 wells/cm 2 , 5000 wells/cm 2 , 1000 wells/cm 2 , 500 wells/cm 2 , or 100 wells/cm 2.
  • the interior surface of the microwells comprises a hydrophilic material that preferably accommodates an aqueous sample; in some cases, the region between the microwells is composed of a hydrophobic material that may preferentially attract a hydrophobic sealing fluid described herein.
  • microwell arrays e.g., Figure 2B may be arranged within a single device.
  • discrete microwell array slides may be arrayed in parallel on a plate holder.
  • at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 25, 50 or 100 microwell arrays are arrayed in parallel.
  • at most 100, 50, 25, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 devices are arrayed in parallel.
  • microwell arrays within a common device may be manipulated simultaneously or sequentially.
  • arrayed devices may be loaded with samples or capsules simultaneously or sequentially.
  • the microwell array may comprise any of a number of different fluids including aqueous, non-aqueous, oils, and organic solvents, such as alcohols.
  • the fluid is used to carry a component, e.g., reagent, microcapsule, or analyte, to a target location such as microwells, output port, etc.
  • the fluid is used to flush the system.
  • the fluid may be used to seal the microwells.
  • any fluid or buffer that is physiologically compatible with the analytes (e.g., cells, molecules) or reagents used in the device may be used.
  • the fluid is aqueous (buffered or not buffered).
  • a sample comprising a population of cells suspended in a buffered aqueous solution may be introduced into the microwell array, allowed to flow through the device, and distributed to the microwells.
  • the fluid passing through the device is nonaqueous (e.g., oil).
  • nonaqueous fluids include but are not limited to: oils, non-polar solvent, hydrocarbon oil, decane (e.g., tetradecane or hexadecane), fluorocarbon oil, fluorinated oil, silicone oil, mineral oil, or other oil.
  • the microcapsules are suspended in a fluid that is compatible with the components of the shell of the microcapsule.
  • Fluids including but not limited to water, alcohols, hydrocarbon oils or fluorocarbon oils are particularly useful fluids for suspending and flowing microcapsules through the microarray device.
  • a sealing fluid may be used to further partition or isolate them within the microwells.
  • the sealing fluid may also be used to seal the individual wells.
  • the sealing fluid may be introduced through the same inlet port that was used to introduce the analyte, reagents and/or microcapsules. But in some cases, the sealing fluid is introduced to the device by a separate inlet port, or through multiple separate inlet ports.
  • the sealing fluid is a non-aqueous fluid (e.g., oil).
  • aqueous solution e.g., solution comprising analytes, free reagents and/or microcapsules
  • the wells themselves, as described herein, may comprise a hydrophilic material that enables wicking of the aqueous fluids (e.g., sample fluid, microcapsule fluid) into individual wells.
  • regions external to the wells comprise hydrophobic material, again to encourage the positioning of the aqueous fluid into the interior of the microwells.
  • the sealing fluid may either remain in the device or be removed.
  • the sealing fluid may be removed, e.g., by flowing through the outlet port.
  • the sealing oil may comprise a volatile oil that can be removed by the application of heat.
  • a fluid may be selected such that its density is equal to, greater than or less than the density of the microcapsules.
  • the microcapsules may be denser than the sealing oil and/or aqueous fluid of the sample and reagents, thereby enabling the microcapsules to remain in the microwells as the sealing oil flows through the device.
  • the capsules may be less dense than the aqueous fluid of the sample or the fluid that the microcapsules are suspended in, as described herein, thereby facilitating movement and distribution of the capsules across the plurality of microwells in a device.
  • a magnetic field may be used to load or direct the capsules into the microwells.
  • a magnetic field may also be used to retain such microcapsules within the wells while the wells are being filled with sample, reagent, and/or sealing fluids.
  • the magnetic field may also be used to remove capsule shells from the wells, particularly following rupture of the capsules.
  • the sealing fluid may remain in the microwells when operations or reactions are conducted therein.
  • the presence of the sealing fluid may act to further partition, isolate, or seal the individual microwells.
  • the sealing fluid may act as a carrier for the microcapsules.
  • sealing fluid comprising
  • microcapsules may be introduced to the device to facilitate distribution of the
  • the sealing fluid may be denser than the microcapsules in order to encourage more even distribution of the microcapsules to the microwells.
  • the microcapsules within the sealing fluid may release reagents to the microwell.
  • the sealing fluid may comprise a chemical or other agent capable of traveling from the sealing fluid to a well (e.g., by leaching or other mechanism) and triggering capsule rupture, where the capsule is present within the microwell or within the sealing fluid.
  • sealing fluids may also be used to seal the microwells following the loading of the analyte, free reagents, and/or microcapsules.
  • the microwells may be sealed with a laminate, tape, plastic cover, oils, waxes, or other suitable material to create an enclosed reaction chamber.
  • the sealants described herein may protect the contents of the microwells from evaporation or other unintended consequences of the reactions or operations. Prevention of evaporation may be particularly necessary when heat is applied to the device, e.g., when heat is applied to stimulate microcapsule release.
  • the laminate seal may also allow recovery of contents from individual wells.
  • a single well of interest may be unsealed (e.g., by removal of the laminate seal) at a given time in order to enable further analysis of an analyte such as by MALDI mass spectrometry.
  • Such applications may be useful in a number of settings, including high-throughput drug screening.
  • analytes, free reagents, and/or microcapsules may be loaded into the present device in any appropriate manner or order.
  • the loading may be random or non-random.
  • a precise number of analytes and/or microcapsules are loaded into each individual microwell.
  • a precise number of analytes and/or microcapsules are loaded into a particular subset of microwells in the plate.
  • an average number of analytes and/or micrcocapsules are loaded into each individual microwell.
  • dry as described herein, in some cases, "dry"
  • microcapsules are loaded into the device, while in other cases “wet” microcapsules are loaded into the device. In some cases, a combination of "dry” and “wet” microcapsules and/or reagents are loaded into the device, either simultaneously or sequentially.
  • the loading of the device may occur in any order and may occur in multiple stages.
  • the microcapsules are pre-loaded into the device, prior to the loading of the analyte.
  • the microcapsules and analyte are loaded concurrently.
  • the analytes are loaded before the microcapsules are loaded.
  • microcapsules and/or analytes may be loaded in multiple stages or multiple times.
  • microcapsules may be loaded into the device both prior to and after analytes are loaded into the device.
  • the microcapsules that are pre-loaded e.g., loaded prior to the analyte introduction
  • the pre-loaded microcapsules contain reagents that are different from the reagents within the microcapsules loaded after analyte introduction.
  • at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 different sets of microcapsules are loaded onto the device.
  • microcapsules are loaded sequentially; or, different sets of microcapsules may also be loaded simultaneously.
  • multiple sets of analytes can be loaded into the device. In some cases, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 different sets of analytes are loaded onto the device. In some cases, the different sets of analytes are loaded sequentially; or, different sets of analytes may also be loaded simultaneously.
  • This disclosure provides devices comprising certain numbers of microcapsules and/or analytes loaded per well. In some cases, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 75, or 100 microcapsules and/or analytes are loaded into each individual microwell. In some cases, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 75, or 100 microcapsules and/or analytes are loaded into each individual microwell. In some cases, on average, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 75, or 100 microcapsules and/or analytes are loaded into each individual microwell.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 75, or 100 microcapsules and/or analytes are loaded into each individual microwell. In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 75, or 100 microcapsules and/or analytes are loaded into each individual micro well.
  • Analytes and/or microcapsules may be applied in a quantity that allows a desired number of analytes to be deposited into an individual microwell. For example, terminal dilution of analytes, such as cells, may achieve the loading of one cell per one microwell or any desired number of analytes per microwell. In some cases, a Poisson distribution is used to direct or predict the final concentration of analytes or microcapsules per well.
  • the microcapsules may be loaded into the microarray device in a particular pattern.
  • certain sections of the device may comprise microcapsules containing a particular reagent (e.g., unique bar-code, enzyme, antibody, antibody subclass, etc.), while other sections of the device may comprise microcapsules containing a different reagent (e.g., a different bar-code, different enzyme, different antibody different antibody subclass, etc.).
  • the microcapsules in one section of the array may contain control reagents. For example, they may contain positive controls that include a control analyte and necessary materials for a reaction.
  • the microcapsules contain negative control reagents such as deactivated enzyme, or a synthetic oligonucleotide sequence that is resistant to fragmentation.
  • negative control reagents may control for the specificity of the sample preparation reaction etc.
  • the negative control microcapsules may comprise the same reagents present in other microcapsules except that the negative control microcapsule may lack a certain reagent (e.g., lysis buffer, polymerase, etc.).
  • the analytes/sample also may be loaded into the microarray device in a particular pattern.
  • certain sections of the device may comprise particular analytes, such as control analytes or analytes deriving from a particular source. This may be used in combination with specific loading of bar codes into known well locations. This feature may allow mapping of specific locations on the array to sequence data, thereby reducing the number of bar codes to be used for labeling reactions.
  • an analyte and reagents may be combined within the droplet with the aid of a microfluidic device.
  • a droplet may be generated that comprises a gel bead (e.g., comprising an oligonucleotide barcode) a nucleic acid analyte, and any other desired reagents.
  • the gel bead, nucleic acid analyte, and reagents in an aqueous phase may be combined at a junction of two or more channels of a microfluidic device.
  • a droplet comprising the resulting mixture may be generated by contacting the aqueous mixture of reagents, gel bead, and nucleic acid analyte with an oil continuous phase.
  • a microcapsule is degradable.
  • the trigger may cause disruption or degradation of the shell or membrane enveloping the microcapsule, disruption or degradation of the interior of a microcapsule, and/or disruption or degradation of any chemical bonds that immobilize a reagent to the microcapsule.
  • exemplary triggers include but are not limited to: chemical triggers, bulk changes, biological triggers, light triggers, thermal triggers, magnetic triggers, and any combination thereof. See, e.g., Esser-Kahn et ah, (2011)
  • Numerous chemical triggers may be used to trigger the disruption or degradation of the microcapsules.
  • Examples of these chemical changes may include, but are not limited to pH-mediated changes to the shell wall, disintegration of the shell wall via chemical cleavage of crosslink bonds, triggered depolymerization of the shell wall, and shell wall switching reactions. Bulk changes may also be used to trigger disruption of the microcapsules.
  • a change in pH of the solution may trigger disruption via a number of different mechanisms.
  • the addition of acid may cause degradation or disassembly of the shell wall through a variety of mechanisms.
  • Addition of protons may disassemble cross-linking of polymers in the shell wall, disrupt ionic or hydrogen bonds in the shell wall, or create nanopores in the shell wall to allow the inner contents to leak through to the exterior.
  • the microcapsule comprises acid-degradable chemical cross-linkers such a ketals.
  • a decrease in pH, particular to a pH lower than 5, may induce the ketal to convert to a ketone and two alcohols and facilitate disruption of the microcapsule.
  • the microcapsules may comprise one or more polyelectrolytes (e.g., PAA, PAAm, PSS, etc.) that are pH sensitive. A decrease in pH may disrupt the ionic- or hydrogen-bonding interactions of such microcapsules, or create nanopores therein.
  • microcapsules comprising polyelectrolytes comprise a charged, gel-based core that expands and contracts upon a change of pH.
  • cross-linkers e.g., disulfide bonds
  • various chemicals can be added to a solution of microcapsules that induce either oxidation, reduction or other chemical changes to polymer components of the shell wall.
  • a reducing agent such as beta-mercaptoethanol, dithiotheritol (DTT), or 2- tris(2- carboxyethyl) phosphine (TCEP) is added such that disulfide bonds in a microcapsule shell wall are disrupted.
  • DTT dithiotheritol
  • TCEP 2- tris(2- carboxyethyl) phosphine
  • enzymes may be added to cleave peptide bonds within the microcapsules, thereby resulting in cleavage of shell wall cross linkers.
  • Depolymerization can also be used to disrupt the microcapsules.
  • a chemical trigger may be added to facilitate the removal of a protecting head group.
  • the trigger may cause removal of a head group of a carbonate ester or carbamate within a polymer, which in turn causes depolymerization and release of reagents from the inside of the capsule.
  • Shell wall switching reactions may be due to any structural change to the porosity of the shell wall.
  • the porosity of a shell wall may be modified, for example, by the addition of azo dyes or viologen derivatives. Addition of energy (e.g., electricity, light) may also be used to stimulate a change in porosity.
  • a chemical trigger may comprise an osmotic trigger, whereby a change in ion or solute concentration of microcapsule solution induces swelling of the capsule. Swelling may cause a buildup of internal pressure such that the capsule ruptures to release its contents.
  • Biological stimuli may also be used to trigger disruption or degradation of microcapsules.
  • biological triggers resemble chemical triggers, but many examples use biomolecules, or molecules commonly found in living systems such as enzymes, peptides, saccharides, fatty acids, nucleic acids and the like.
  • microcapsules may comprise polymers with peptide cross-links that are sensitive to cleavage by specific proteases. More specifically, one example may comprise a microcapsule comprising GFLGK peptide cross links.
  • a biological trigger such as the protease Cathepsin B, the peptide cross links of the shell well are cleaved and the contents of the capsule are released.
  • microcapsules comprise a shell wall comprising cellulose. Addition of the hydro lytic enzyme chitosan serves as biologic trigger for cleavage of cellulosic bonds, depolymerization of the shell wall, and release of its inner contents.
  • the microcapsules may also be induced to release their contents upon the application of a thermal stimulus.
  • a change in temperature can cause a variety changes to the microcapsule.
  • a change in heat may cause melting of a microcapsule such that the shell wall disintegrates.
  • the heat may increase the internal pressure of the inner components of the capsule such that the capsule ruptures or explodes.
  • the heat may transform the capsule into a shrunken dehydrated state.
  • the heat may also act upon heat-sensitive polymers within the shell of a microcapsule to cause disruption of the microcapsule.
  • a microcapsule comprises a thermo-sensitive hydrogel shell encapsulating one or more emulsified reagent particles.
  • the hydrogel material of the outer shell wall shrinks.
  • the sudden shrinkage of the shell ruptures the capsule and allows the reagents of the inside of the capsule to squirt out in the sample preparation solution in the microwell.
  • the shell wall may comprise a diblock polymer, or a mixture of two polymers, with different heat sensitivities.
  • One polymer may be particularly likely to shrink after the application of heat, while the other is more heat-stable.
  • the heat-sensitive polymer may shrink, while the other remains intact, causing a pore to form.
  • a shell wall may comprise magnetic nanoparticles. Exposure to a magnetic field may cause the generation of heat, leading to rupture of the microcapsule.
  • a device of this disclosure may comprise magnetic particles for either purpose.
  • incorporation of Fe304 nanoparticles into polyelectrolyte containing capsules triggers rupture in the presence of an oscillating magnetic field stimulus.
  • a microcapsule may also be disrupted or degraded as the result of electrical stimulation. Similar to magnetic particles described in the previous section, electrically sensitive particles can allow for both triggered rupture of the capsules as well as other functions such as alignment in an electric field, electrical conductivity or redox reactions. In one example, microcapsules containing electrically sensitive material are aligned in an electric field such that release of inner reagents can be controlled. In other examples, electrical fields may induce redox reactions within the shell wall itself that may increase porosity.
  • a light stimulus may also be used to disrupt the microcapsules.
  • Numerous light triggers are possible and may include systems that use various molecules such as nanoparticles and chromophores capable of absorbing photons of specific ranges of wavelengths.
  • metal oxide coatings can be used as capsule triggers.
  • UV irradiation of polyelectrolyte capsules coated with Si02/Ti02 may result in disintegration of the capsule wall.
  • photo switchable materials such as azobenzene groups may be incorporated in the shell wall.
  • chemicals such as these undergo a reversible cis-to-trans isomerization upon absorption of photons.
  • incorporation of photo switches result in a shell wall that may disintegrate or become more porous upon the application of a light trigger.
  • a device of this disclosure may be used in combination with any apparatus or device that provides such trigger or stimulus.
  • the stimulus is thermal
  • a device may be used in combination with a heated or thermally controlled plate, which allows heating of the micro wells and may induce the rupture of capsules.
  • Any of a number of heat transfers may be used for thermal stimuli, including but not limited to applying heat by radiative heat transfer, convective heat transfer, or conductive heat transfer.
  • the stimulus is a biological enzyme
  • the enzyme may be injected into a device such that it is deposited into each microwell.
  • a device may be used in combination with a magnetic or electric plate.
  • a chemical stimulus may be added to a partition and may exert its function at various times after contacting a chemical stimulus with a microcapsule.
  • the speed at which a chemical stimulus exerts its effect may vary depending on, for example, the amount/concentration of a chemical stimulus contacted with a microcapsule and/or the particular chemical stimulus used.
  • a droplet may comprise a degradable gel bead (e.g., a gel bead comprising chemical cross-linkers, such as, for example, disulfide bonds).
  • a chemical stimulus e.g., a reducing agent
  • a chemical stimulus e.g., a reducing agent
  • the chemical stimulus may degrade the gel bead immediately on contact with the gel bead, soon after (e.g., about 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 min) contact with the gel bead, or at a later time.
  • degradation of the gel bead may occur before, during, or after a further processing step, such as, for example, a thermal cycling step as described herein.
  • the sample preparation reaction may proceed in a device. Reactions within a device may be incubated for various periods of times depending on the reagents used in the sample reactions.
  • a device may also be used in combination with other devices that aid in the sample preparation reaction.
  • a device may be used in combination with a PCR thermocycler.
  • a thermocycler may comprise a plurality of wells. In cases where partitions are droplets, the droplets may be entered into the wells of the thermocycler. In some cases, each well may comprise multiple droplets, such that when thermal cycling is initiated, multiple droplets are thermal cycled in each well.
  • a device may be used in combination with a shaking apparatus.
  • a device may utilize a method comprising the application of liquid or gas to flush out the contents of the individual microwells.
  • the liquid comprises an immiscible carrier fluid that preferentially wets the microwell array material. It may also be immiscible with water so as to flush the reaction products out of the well.
  • the liquid may be an aqueous fluid that can be used to flush out the samples out of the wells. After flushing of the contents of the microwells, the contents of the microwells are pooled for a variety of downstream analyses and applications. VII. Applications
  • Figure 4A provides a general flow of many of the methods of the present disclosure; and Figure 4B provides a generally annotated version of 4A.
  • One or more microcapsule(s) that contain reagents 410 may be pre-loaded into microwells, followed by addition of an analyte, which, in this particular Figure, is a nucleic acid analyte 420.
  • the microwells may then be sealed 430 by any method, such as by application of a sealing fluid.
  • the inlet and outlet ports may also be sealed, for example to prevent evaporation.
  • a stimulus e.g., heat, chemical, biological, etc.
  • a stimulus e.g., heat, chemical, biological, etc.
  • an incubation step 440 may occur in order to enable the reagents perform a particular function such as lysis of cells, digestion of protein, fragmentation of high molecular weight nucleic acids, or ligation of oligonucleotide bar codes.
  • the contents of the microwells may be recovered either singly or in bulk.
  • a device of this disclosure may have a wide variety of uses in the manipulation, preparation, identification and/or quantification of analytes.
  • the analyte is a cell or population of cells.
  • the population of cells may be homogeneous (e.g., from a cell line, of the same cell type, from the same type of tissue, from the same organ, etc.) or heterogenous (mixture of different types of cells).
  • the cells may be primary cells, cell lines, recombinant cells, primary cells, encapsulated cells, free cells, etc.
  • the analytes may also be molecules, including but not limited to: polypeptides, proteins, antibodies, enzymes, nucleic acids, saccharides, small molecules, drugs, and the like.
  • nucleic acids include but are not limited to: DNA, R A, dNTPs, ddNTPs, amplicons, synthetic nucleotides, synthetic polynucleotides, polynucleotides, oligonucleotides, peptide nucleic acids, cDNA, dsDNA, ssDNA, plasmid DNA, cosmid DNA, high Molecular Weight (MW) DNA, chromosomal DNA, genomic DNA, viral DNA, bacterial DNA, mtDNA (mitochondrial DNA), mRNA, rRNA, tRNA, nRNA, siRNA, snRNA, snoRNA, scaRNA, microRNA, dsRNA, ribozyme, riboswitch and viral RNA (e.g., retro
  • the analytes are pre -mixed with one or more additional materials, such as one or more reagents (e.g., ligase, protease, polymerase) prior to being loaded into the device.
  • one or more reagents e.g., ligase, protease, polymerase
  • the analytes are pre -mixed with microcapsules comprising one or more reagents prior to being loaded onto the device.
  • the samples may be derived from a variety of sources including human, mammal, non-human mammal, ape, monkey, chimpanzee, plant, reptilian, amphibian, avian, fungal, viral or bacterial sources.
  • Samples such as cells, nucleic acids and proteins may also be obtained from a variety of clinical sources such as biopsies, aspirates, blood draws, urine samples, formalin fixed embedded tissues and the like.
  • a device of this disclosure may also enable the analytes to be tagged or tracked in order to permit subsequent identification of an origin of the analytes. This feature is in contrast with other methods that use pooled or multiplex reactions and that only provide measurements or analyses as an average of multiple samples.
  • the physical partitioning and assignment of a unique identifier to individual analytes allows acquisition of data from individual samples and is not limited to averages of samples.
  • nucleic acids or other molecules derived from a single cell may share a common tag or identifier and therefore may be later identified as being derived from that cell. Similarly, all of the fragments from a single strand of nucleic acid may be tagged with the same identifier or tag, thereby permitting subsequent
  • gene expression products e.g., mR A, protein
  • the device can be used as a PCR amplification control. In such cases, multiple amplification products from a PCR reaction can be tagged with the same tag or identifier. If the products are later sequenced and demonstrate sequence differences, differences among products with the same identifier can then be attributed to PCR error.
  • the analytes may be loaded onto the device before, after, or during loading of the microcapsules and/or free reagents.
  • the analytes are encapsulated into microcapsules before loading into the microcapsule array.
  • nucleic acid analytes may be encapsulated into a microcapsule, which is then loaded onto the device and later triggered to release the analytes into an appropriate microwell.
  • Any analytes such as DNA or cells, may be loaded in solution or as analytes encapsulated in a capsule.
  • homogeneous or heterogeneous populations of molecules e.g., nucleic acids, proteins, etc.
  • homogeneous or heterogeneous populations of cells are encapsulated into microcapsules and loaded onto the device.
  • the microcapsules may comprise a random or specified number of cells and/or molecules.
  • the microcapsules may comprise no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30,40 ,50, 60, 70, 80, 90, 100, 500, 1000, 5000, or 10000 cells and/or molecules per microcapsule.
  • the microcapsules comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30,40, 50, 60, 70, 80, 90, 100, 500, 1000, 5000, or 10000 cells and/or molecules per
  • Fluidic techniques and any other techniques may be used to encapsulate the cells and/or molecules into the microcapsules.
  • sequences provided herein are useful for preparation of an analyte prior to a down-stream application such as a sequencing reaction.
  • a sequencing method is classic Sanger sequencing.
  • Sequencing methods may include, but are not limited to: high-throughput sequencing, pyrosequencing, sequencing-by-synthesis, single-molecule sequencing, nanopore sequencing, sequencing- by-ligation, sequencing-by-hybridization, RNA-Seq (Illumina), Digital Gene Expression (Helicos), Next generation sequencing, Single Molecule Sequencing by Synthesis (SMSS)(Helicos), massively-parallel sequencing, Clonal Single Molecule Array (Solexa), shotgun sequencing, Maxim-Gilbert sequencing, primer walking, and any other sequencing methods known in the art.
  • the devices disclosed herein may be used in applications that involve the assignment of unique identifiers, or molecular bar codes, to analytes.
  • the unique identifier is a bar-code oligonucleotide that is used to tag the analytes; but, in some cases, different unique identifiers are used.
  • the unique identifier is an antibody, in which case the attachment may comprise a binding reaction between the antibody and the analyte (e.g., antibody and cell, antibody and protein, antibody and nucleic acid).
  • the unique identifier is a dye, in which case the attachment may comprise intercalation of the dye into the analyte molecule (such as intercalation into DNA or RNA) or binding to a probe labeled with the dye.
  • the unique identifier may be a nucleic acid probe, in which case the attachment to the analyte may comprise a hybridization reaction between the nucleic acid and the analyte.
  • the reaction may comprise a chemical linkage between the identifier and the analyte.
  • the reaction may comprise addition of a metal isotope, either directly to the analyte or by a probe labeled with the isotope.
  • the method comprises attaching oligonucleotide bar codes to nucleic acid analytes through an enzymatic reaction such as a ligation reaction.
  • the ligase enzyme may covalently attach a DNA bar code to fragmented DNA (e.g., high molecular-weight DNA).
  • fragmented DNA e.g., high molecular-weight DNA
  • oligonucleotide primers containing bar code sequences may be used in amplification reactions (e.g., PCR, qPCR, reverse-transcriptase PCR, digital PCR, etc.) of the DNA template analytes, thereby producing tagged analytes.
  • amplification reactions e.g., PCR, qPCR, reverse-transcriptase PCR, digital PCR, etc.
  • the contents of individual microwells may be recovered via the outlet port in the device for further analyses.
  • the unique identifiers may be introduced to the device randomly or nonrandomly. In some cases, they are introduced at an expected ratio of unique identifiers to microwells. For example, the unique identifiers may be loaded so that more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers are loaded per microwell. In some cases, the unique identifiers may be loaded so that less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers are loaded per microwell.
  • the average number of unique identifiers loaded per microwell is less than, or greater than, about 0.0001, 0.001, 0.01, 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers per microwell.
  • the unique identifiers also may be loaded so that a set of one or more identical identifiers are introduced to a particular well. Such sets may also be loaded so that each microwell contains a different set of identifiers. For example, a population of
  • microcapsules may be prepared such that a first microcapsule in the population comprises multiple copies of identical unique identifiers (e.g., nucleic acid bar codes, etc.) and a second microcapsule in the population comprises multiple copies of a unique identifier that differs from within the first microcapsule.
  • the population of microcapsules may comprise multiple microcapsules (e.g., greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000, 5000, 10000, 100000, 1000000, 10000000, 100000000, or 1000000000 microcapsules), each containing multiple copies of a unique identifier that differs from that contained in the other microcapsules.
  • the population may comprise greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000, 5000, 10000, 100000, 1000000, 10000000, 100000000, or 1000000000 microcapsules with identical sets of unique identifiers. In some cases, the population may comprise greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000, 5000, 10000, 100000, 1000000, 10000000, 100000000, or
  • microcapsules wherein the microcapsules each comprise a different combination of unique identifiers.
  • the different combinations overlap, such that a first microcapsule may comprise, e.g., unique identifiers A, B, and C, while a second microcapsule may comprise unique identifiers A, B, and D.
  • the different combinations do not overlap, such that a first microcapsule may comprise, e.g., unique identifiers A, B, and C, while a second microcapsule may comprise unique identifiers D, E, and F.
  • the unique identifiers may be loaded into the device at an expected or predicted ratio of unique identifiers per analyte (e.g., strand of nucleic acid, fragment of nucleic acid, protein, cell, etc.) In some cases, the unique identifiers are loaded in the microwells so that more than about 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers are loaded per individual analyte in the microwell.
  • analyte e.g., strand of nucleic acid, fragment of nucleic acid, protein, cell, etc.
  • the unique identifiers are loaded in the microwells so that less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers are loaded per individual analyte in the microwell.
  • the average number of unique identifiers loaded per analyte is less than, or greater than, about 0.0001, 0.001, 0.01, 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 500, 1000, 5000, 10000, or 200000 unique identifiers per analyte.
  • such identifiers may be copies of the same identifier, or multiple different identifiers.
  • the attachment process may be designed to attach multiple identical identifiers to a single analyte, or multiple different identifiers to the analyte.
  • the unique identifiers may be used to tag a wide range of analytes, including cells or molecules.
  • unique identifiers e.g., bar code oligonucleotides
  • the unique identifiers may also bind to cells, include the external surface of a cell, a marker expressed on the cell or components within the cell such as organelles, gene expression products, genomic DNA, mitochondrial DNA, RNA, mRNA, or proteins.
  • the unique identifiers also may be designed to bind or hybridize nucleic acids (e.g., DNA, RNA) present in permeabilized cells, which may or may not be otherwise intact.
  • the unique identifiers may be loaded onto the device either singly or in combination with other elements (e.g., reagents, analytes). In some cases, free unique identifiers are pooled with the analytes and the mixture is loaded into the device. In some cases, unique identifiers encapsulated in microcapsules are pooled with the analytes, prior to loading of the mixture onto the device. In still other cases, free unique identifiers are loaded into the microwells prior to, during (e.g., by separate inlet port), or following the loading of the analytes. In still other cases, unique identifiers encapsulated in
  • microcapsules are loaded into the microwells prior to, concurrently with (e.g., by separate inlet port), or after loading of the analytes.
  • analytes may be important to determine whether individual analytes each receive a different unique identifier (e.g., oligonucleotide bar code). If the population of unique identifiers introduced into the device is not significantly diverse, different analytes may possibly be tagged with identical identifiers.
  • the devices disclosed herein may enable detection of analytes tagged with the same identifier.
  • a reference analyte may be included with the population of analytes introduced into the device.
  • the reference analyte may be, for example, a nucleic acid with a known sequence and a known quantity.
  • unique identifiers may be attached to the analytes, as described herein. If the unique identifiers are oligonucleotide bar codes and the analytes are nucleic acids, the tagged analytes may subsequently be sequenced and quantified. These methods may indicate if one or more fragments and/or analytes may have been assigned an identical bar code.
  • a method disclosed herein may comprise loading the device with the reagents necessary for the assignment of bar codes to the analytes.
  • reagents including, but not limited to, ligase enzyme, buffer, adapter oligonucleotides, a plurality of unique identifier DNA bar codes and the like may be loaded into the device.
  • reagents including but not limited to a plurality of PCR primers, oligonucleotides containing unique identifying sequence, or bar code sequence, DNA polymerase, DNTPs, and buffer and the like may be loaded into the device.
  • the reagents may be loaded as free reagents or as reagents encapsulated in microcapsules.
  • Nucleic acid sequencing may begin with the physical partitioning of sample analytes into microwells at a particular density (e.g., about 1 analyte per microwell or other density described herein).
  • a density e.g., about 1 analyte per microwell or other density described herein.
  • the devices provided herein may be used to prepare analytes (e.g., nucleic acid analytes) in such a manner that enables phasing or linkage information to be subsequently obtained. Such information may allow for the detection of linked genetic variations in sequences, including genetic variations (e.g., SNPs, mutations, indels, copy number variations, transversions, translocations, inversions, etc.) that are separated by long stretches of nucleic acids. These variations may exist in either a cis or trans
  • cis relationships two or more genetic variations may exist in the same polynucleic acid molecule or strand.
  • trans relationships two or more genetic variations may exist on multiple nucleic acid molecules or strands.
  • a method of determining nucleic acid phasing may comprise loading a nucleic acid sample (e.g., a nucleic acid sample that spans a given locus or loci) into a device disclosed herein, distributing the sample such that at most one molecule of nucleic acid is present per microwell, and fragmenting the sample within the microwells.
  • the method may further comprise attaching unique identifiers (e.g., bar codes) to the fragmented nucleic acids as described herein, recovering the nucleic acids in bulk, and performing a subsequent sequencing reaction on the samples in order to detect genetic variations, such as two different genetic variations.
  • phase information may be important for the characterization of the analyte, particularly if the analyte derives from a subject at risk of, having, or suspected of a having a particular disease or disorder (e.g., hereditary recessive disease such as Cystic Fibrosis, cancer, etc.).
  • a particular disease or disorder e.g., hereditary recessive disease such as Cystic Fibrosis, cancer, etc.
  • the information may be able to distinguish between the following possibilities: (1) two genetic variations within the same gene on the same strand of DNA and (2) two genetic variations within the same gene but located on separate strands of DNA. Possibility (1) may indicate that one copy of the gene is normal and the individual is free of the disease, while possibility (2) may indicate that the individual has or will develop the disease, particularly if the two genetic variations are damaging to the function of the gene when present within the same gene copy. Similarly, the phasing information may also be able to distinguish between the following possibilities: (1) two genetic variations, each within a different gene on the same strand of DNA and (2) two genetic variations, each within a different gene but located on separate strands of DNA.
  • the devices provided herein may be used to prepare cellular analytes in such a manner that enables cell-specific information to be subsequently obtained. Such information may enable detection of genetic variations (e.g., SNPs, mutations, indels, copy number variations, transversions, translocations, inversions, etc.) on a cell-by-cell basis, thereby enabling a determination of whether the genetic variation(s) are present in the same cell or two different cells.
  • genetic variations e.g., SNPs, mutations, indels, copy number variations, transversions, translocations, inversions, etc.
  • a method of determining nucleic acid cell-specific information may comprise loading a cellular sample (e.g., a cellular sample from a subject) into a device disclosed herein, distributing the sample such that at most one cell is present per microwell, lysing the cells, and then tagging the nucleic acids within the cells with unique identifiers using a method described herein.
  • a cellular sample e.g., a cellular sample from a subject
  • microcapsules comprising unique identifiers are loaded in the microwell array device (either before, during, or after the loading of the cellular analytes) in such a manner that each cell is contacted with a different
  • nucleic acids with identical unique identifiers may be determined to originate from the same cell, while nucleic acids with different unique identifiers may be determined to originate from different cells.
  • the methods herein may be used to detect the distribution of oncogenic mutations across a population of cancer tumor cells.
  • some of the cells may have a mutation, or amplification, of an oncogene (e.g., HER2, BRAF, EGFR, KRAS) on two strands of DNA (homozygous), while others may be heterozygous for the mutation, while still other cells may be wild-type and comprise no mutations or other variation in the oncogene.
  • the methods described herein may be able to detect these differences, and also may enable quantification of the relative numbers of homozygous, heterozygous, and wild-type cells. Such information may be used to stage a particular cancer or to monitor the progression of the cancer over time.
  • this disclosure provides methods of identifying mutations in two different oncogenes (e.g., KRAS and EGFR). If the same cell comprises genes with both mutations, this may indicate a more aggressive form of cancer. In contrast, if the mutations are located in two different cells, this may indicate that the cancer is more benign, or less advanced.
  • oncogenes e.g., KRAS and EGFR
  • a plurality of cells such as from a tumor biopsy, is loaded into a device. Single cells from the sample are deposited into individual wells and labeled with a DNA bar code.
  • Loading of cells into a device may be achieved through non-random loading.
  • Parameters for non-random loading of analytes, such as cells, may be understood using an interference function such that: the well (measure of interfer
  • cells may be lysed and many subsequent reactions are possible, including RNA amplification, DNA amplification or antibody screening for different target proteins and genes in individual cells.
  • the contents of the cells may be pooled together and could be further analyzed, such as by DNA sequencing.
  • further analyses may be possible including but not limited to quantification of different gene levels or nucleic acid sequencing of individual cells.
  • it may be determined whether the tumor comprises cells with different genetic backgrounds (e.g., cancer clones and subclones). The relative number of each type of cell may also be calculated.
  • the device can be used for purposes of controlling for amplification errors, such as PCR errors.
  • a nucleic acid sample may be partitioned into the microwells of the device. Following partitioning, the sample may be subjected to a PCR amplification reaction within the microwells.
  • the PCR products within a microwell may be tagged with the same unique identifier, using a method described herein. If the products are later sequenced and demonstrate sequence differences, differences among products with the same identifier can then be attributed to PCR error.
  • a device may be used to detect gene product (e.g., protein, mRNA) expression levels in a sample, often on a cell-by-cell basis.
  • a sample may comprise individual cells, a pool of mRNA extract from cells, or other collection of gene products.
  • single cells may be loaded into microwells.
  • a pool of mRNA or other gene product may be loaded such that a desired quantity of mRNA molecules is loaded into individual microwells.
  • the methods provided herein may be particularly useful for RNA analysis.
  • unique identifiers may be assigned to mRNA analytes either directly or to cDNA products of a reverse transcription reaction performed on the mRNA analytes.
  • the reverse transcription reaction may be conducted within the microwells of the device following loading of the analytes.
  • Reagents for the reaction may include but are not limited to reverse transcriptase, DNA polymerase enzyme, buffer, dNTPs, oligonucleotide primers, oligonucleotide primers containing bar code sequences and the like.
  • One or more reagents may be loaded into microcapsules or loaded freely in solution into the device or a combination thereof.
  • Sample preparation may then be conducted, such as by fragmenting the cDNA and attaching unique identifiers to the fragments.
  • the nucleic acid products of the reaction may be further analyzed, such as by sequencing.
  • a device may be used to characterize multiple cell markers, similar to a flow cytometer.
  • Any cell marker may be characterized, including cell-surface markers (e.g., extracellular proteins, transmembrane markers) and markers located within the internal portion of a cell (e.g., RNA, mRNA, microRNA, multiple copies of genes, proteins, alternative splicing products, etc.).
  • cell-surface markers e.g., extracellular proteins, transmembrane markers
  • markers located within the internal portion of a cell e.g., RNA, mRNA, microRNA, multiple copies of genes, proteins, alternative splicing products, etc.
  • cells may be partitioned within the device, as described herein, so that at most one cell is present within a microwell.
  • Cell markers such as nucleic acids (e.g., R A) may be extracted and/or fragmented prior to being labeled with a unique identifier (e.g., molecular bar code).
  • the nucleic acids may be labeled with a unique identifier without being extracted and/or fragmented.
  • the nucleic acids may then be subjected to further analysis such as sequencing reactions designed to detect multiple gene expression products.
  • Such analysis may be useful in a number of fields. For example, if the starting cells are immune cells (e.g., T cells, B cells, macrophages, etc.), the analysis may provide information regarding multiple expressed markers and enable immunophenotyping of the cells, for example by identifying different CD markers of the cells (e.g., CD3, CD4, CD8, CD 19, CD20, CD 56, etc.).
  • CD markers e.g., CD3, CD4, CD8, CD 19, CD20, CD 56, etc.
  • markers can also be used in conjunction with markers that are not necessarily immunophenotyping markers, such as markers of pathogenic infection (e.g., viral or bacterial protein, DNA, or RNA).
  • the device may be used to identify at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 500, 700, 1000, 5000, 10000, 50000, or 100000 different gene expression products or other form of cellular markers on a single-cell basis.
  • such methods do not comprise use of dyes or probes (e.g., fluorescent probes or dyes).
  • Gene expression product analysis may be useful in numerous fields including immunology, cancer biology (e.g., to characterize the existence, type, stage,
  • stem cell biology e.g., in order to characterize the differentiation state of a stem cell, potency of a stem cell, cellular type of a stem cell, or other features of a stem cell
  • microbiology e.g., in order to characterize the differentiation state of a stem cell, potency of a stem cell, cellular type of a stem cell, or other features of a stem cell
  • the gene expression analysis may also be used in drug screening applications, for example to evaluate the effect of a particular drug or agent on the gene expression profile of particular cells.
  • Ranges can be expressed herein as from “about” one particular value, and/or to "about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. The term "about” as used herein refers to a range that is 15% plus or minus from a stated numerical value within the context of the particular usage. For example, about 10 would include a range from 8.5 to 11.5.
  • microwell array generally refers to a predetermined spatial arrangement of microwells.
  • Microwell array devices that comprise a microcapsule may also be referred to as "microwell capsule arrays.”
  • array may be used herein to refer to multiple arrays arranged on a surface, such as would be the case where a surface has multiple copies of an array. Such surfaces bearing multiple arrays may also be referred to as “multiple arrays” or “repeating arrays.”
  • a microwell capsule array is prepared to perform nucleic acid sequencing on individual human B-cells taken from a blood sample. Approximately 15,000 cells are harvested and used for loading into the device. A device of this disclosure and containing 150,000 microwells is used. Each well is cylindrical in shape having a diameter of 125 um and a height of 125 um, allowing at most 1 capsule to be loaded per well. Microcapsules made through emulsion polymerization with a PNIPAM hydrogel shell wall are created such that the microcapsules have a diameter of 100 um for loading in the device. The microcapsules are created such that the PNIPAM shell contains magnetic iron particles. The outer surface of the shell is then chemically coupled to a antibody specific to a transmembrane B cell receptor on the outside of a B cell.
  • reagents are simultaneously loaded into the capsules.
  • Reagents necessary for cell lysis and labeling individual DNA strands of the cells with DNA barcodes are loaded into capsules.
  • Reagents for cell lysis include a mild non-ionic detergent, buffer and salt.
  • Reagents for the addition of DNA bar codes to genomic DNA included restriction enzymes, ligase, and > 10,000,000 unique DNA oligonucleotides are loaded into capsules. Capsules are designed to be sensitive to rupture at greater than 65 C.
  • Capsules are prepared to be applied to the microcapsule array.
  • the array is placed on a magnetic temperature controlled hot plate.
  • Microcapsules are added to a sample of B cells such that one B cell is able to bind to one capsule.
  • Capsule-cell conjugates are applied in aqueous carrier solution in a quantity in excess to the relative number of wells.
  • gentle pipetting of capsules-cells into the inlet port followed by application of a vacuum manifold to the outlet port distributes the capsules throughout the device.
  • a magnetic field is applied through the plate.
  • Excess capsule-cell solution is removed via pipetting through the outlet port.
  • Each capsule-cell conjugate is trapped and positioned in individual wells via the magnetic field.
  • a carrier oil (or sealing fluid) is applied to the device to remove any excess aqueous solution bridging adjacent micro wells.
  • the carrier oil applied to the inlet and excess oil is recovered at the outlet with a vacuum manifold. After the carrier oil is applied, the inlet and outlet ports are sealed with tape.
  • the device is then heated, via the magnetic temperature controlled hot plate, to a temperature of 70 C for 10 min to allow for capsule rupture and cell lysis.
  • the hot plate is then switched to 37 C, for restriction and ligation, for up to 1 hour.
  • the contents of the wells are recovered.
  • the inlet and outlet ports of the device are unsealed and nitrogen gas is applied to the device to flush out the individual components of the microwells.
  • the sample is collected in bulk via a pipette at the outlet port, while the magnetic field retains ruptured capsule shells in individual microwells.
  • the sample is then sequenced using a multiplex sequencing strategy known in the art. Bar coding of individual cells allows for sequencing information to be gained for individual cells rather than as an average of multiple cells. Based upon the number of cells sequenced and bar codes assigned, SNP cell-specific information is gained.
  • the number of reads for individual bar codes can be counted to provide insight into the distribution of different types of cells with varying genetic backgrounds, within the original population of B cells.
  • a microwell capsule array is prepared to perform nucleic acid sequencing on individual strands of DNA isolated from a population of human skin cells. Cells are lysed using detergent and heat and approximately 15,000 copies of diploid DNA are precipitated via chloroform/ethanol extraction. A resuspension of DNA is loaded into the device with approximately 10,000 copies of haploid DNA.
  • a device of this disclosure, with 300,000 microwells is used. Each well is cylindrical in shape having a diameter of 125 um and a height of 125 um, allowing at most 1 capsule to be loaded per well.
  • Microcapsules made through emulsion polymerization with a PNIPAM hydrogel shell wall are created to a specification of a sphere with a diameter of 100 um for loading into the device.
  • reagents are simultaneously loaded into the capsules.
  • the reagents include reagents necessary for labeling individual DNA strands with DNA barcodes, including restriction enzymes, ligase, and > 10,000,000 unique DNA oligonucleotides. Capsules designed to be sensitive to rupture at greater than 65 C are used for the encapsulation.
  • Capsules are applied aqueous carrier solution in an excess to the relative number of wells. Gentle pipetting of capsules into the inlet followed by application of a vacuum manifold to the outlet distributed the capsules throughout the device. After excess capsule solution is removed, a suspension of DNA in buffer is applied to the device in a similar fashion as the capsules.
  • a carrier oil is applied to the device to remove any excess aqueous solution bridging adjacent microwells.
  • the carrier oil is applied to the inlet port and excess oil is recovered at the outlet port with a vacuum manifold. After the carrier oil is applied, the inlet and outlet ports are sealed with tape.
  • the device is then placed on a temperature controlled hot plate and heated to temperature of 70 C for 10 min to allow for capsule rupture. Reagents are released into the sample preparation reaction.
  • the hot plate is then switched to 37 C, for restriction and ligation, for up to 1 hour.
  • the inlet and outlet ports of the device are unsealed and nitrogen gas is applied to the device to flush out the individual components of the microwells.
  • the sample products, en bulk, are collected via pipette at the outlet port.
  • the sample is then sequenced to sufficient coverage (e.g., 500) using a multiplex sequencing strategy known in the art.
  • Bar coding of individual DNA strands allows for sequencing information to be gained from individual strands rather than as an average of entire sample of DNA. Based upon the number of DNA strands sequenced and bar codes assigned, SNP phasing/haplotyping information is gained and many repetitive regions of DNA can be resolved. In addition, a substantial boost in accuracy can be gained by discarding mutations that appear randomly with respect to haplotypes, as those are likely to be sequencing errors.

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EP13829413.7A EP2885418A4 (en) 2012-08-14 2013-08-13 MICROCAPSE COMPOSITIONS AND METHOD THEREFOR
BR112015003354A BR112015003354A8 (pt) 2012-08-14 2013-08-13 métodos e composições de microcápsula
EP24176310.1A EP4397767A3 (en) 2012-08-14 2013-08-13 Microcapsule compositions and methods
CN202210540740.9A CN114891871A (zh) 2012-08-14 2013-08-13 微胶囊组合物及方法
CA2881685A CA2881685C (en) 2012-08-14 2013-08-13 Microcapsule compositions and methods
CN201380053555.6A CN104769127A (zh) 2012-08-14 2013-08-13 微胶囊组合物及方法
EP21159422.1A EP3901273A1 (en) 2012-08-14 2013-08-13 Microcapsule compositions and methods
JP2015527549A JP6367196B2 (ja) 2012-08-14 2013-08-13 マイクロカプセル組成物および方法
KR1020157006549A KR102090851B1 (ko) 2012-08-14 2013-08-13 마이크로캡슐 조성물 및 방법
MX2015001939A MX364957B (es) 2012-08-14 2013-08-13 Composiciones y metodos para microcapsulas.
IN1126DEN2015 IN2015DN01126A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) 2012-08-14 2013-08-13
AU2013302756A AU2013302756C1 (en) 2012-08-14 2013-08-13 Microcapsule compositions and methods
CN202010761140.6A CN111748607B (zh) 2012-08-14 2013-08-13 微胶囊组合物及方法
US14/316,447 US10221442B2 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
KR1020227005065A KR102436171B1 (ko) 2013-06-27 2014-06-26 샘플 처리를 위한 조성물 및 방법
US14/316,383 US20140378345A1 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
PCT/US2014/044398 WO2014210353A2 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
KR1020227028867A KR102642680B1 (ko) 2013-06-27 2014-06-26 샘플 처리를 위한 조성물 및 방법
EP24158974.6A EP4357493A3 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
BR112015032512A BR112015032512A8 (pt) 2013-06-27 2014-06-26 método de implementação de código de barras de materiais de amostra
AU2014302277A AU2014302277A1 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
US14/316,398 US20150005199A1 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
MX2015016968A MX361481B (es) 2013-06-27 2014-06-26 Composiciones y metodos para procesamiento de muestras.
KR1020167002243A KR102212234B1 (ko) 2013-06-27 2014-06-26 샘플 처리를 위한 조성물 및 방법
CA2915499A CA2915499A1 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
CN201480047858.1A CN105492607B (zh) 2013-06-27 2014-06-26 用于样品处理的组合物和方法
CN202311514025.9A CN117568449A (zh) 2013-06-27 2014-06-26 用于样品处理的组合物和方法
CN201910621839.XA CN110592182B (zh) 2013-06-27 2014-06-26 用于样品处理的组合物和方法
KR1020217002986A KR102366116B1 (ko) 2013-06-27 2014-06-26 샘플 처리를 위한 조성물 및 방법
US14/316,463 US20140378322A1 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
US14/316,431 US20150005200A1 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
US14/316,416 US20140378349A1 (en) 2012-08-14 2014-06-26 Compositions and methods for sample processing
EP14817610.0A EP3013957B2 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
JP2016524208A JP6563912B2 (ja) 2013-06-27 2014-06-26 サンプル処理のための組成物及び方法
EP18189200.1A EP3467160A1 (en) 2013-06-27 2014-06-26 Compositions and methods for sample processing
IL237156A IL237156B (en) 2012-08-14 2015-02-09 Microcapsule preparation and methods
US15/718,764 US20180094298A1 (en) 2012-08-14 2017-09-28 Compositions and methods for sample processing
US15/718,893 US20180051321A1 (en) 2012-08-14 2017-09-28 Compositions and methods for sample processing
US15/887,711 US20180179580A1 (en) 2012-08-14 2018-02-02 Compositions and methods for sample processing
AU2018203012A AU2018203012B2 (en) 2012-08-14 2018-05-01 Microcapsule compositions and methods
US16/052,486 US10323279B2 (en) 2012-08-14 2018-08-01 Methods and systems for processing polynucleotides
US16/056,231 US11591637B2 (en) 2012-08-14 2018-08-06 Compositions and methods for sample processing
US16/231,185 US10400280B2 (en) 2012-08-14 2018-12-21 Methods and systems for processing polynucleotides
US16/242,962 US20190203262A1 (en) 2012-08-14 2019-01-08 Compositions and methods for sample processing
US16/246,322 US10597718B2 (en) 2012-08-14 2019-01-11 Methods and systems for sample processing polynucleotides
US16/294,769 US10450607B2 (en) 2012-08-14 2019-03-06 Methods and systems for processing polynucleotides
US16/395,090 US10669583B2 (en) 2012-08-14 2019-04-25 Method and systems for processing polynucleotides
US16/435,362 US10626458B2 (en) 2012-08-14 2019-06-07 Methods and systems for processing polynucleotides
US16/435,417 US10752950B2 (en) 2012-08-14 2019-06-07 Methods and systems for processing polynucleotides
US16/736,323 US12098423B2 (en) 2012-08-14 2020-01-07 Methods and systems for processing polynucleotides
US16/844,141 US11441179B2 (en) 2012-08-14 2020-04-09 Methods and systems for processing polynucleotides
AU2020203602A AU2020203602C1 (en) 2012-08-14 2020-06-01 Microcapsule compositions and methods
US16/998,425 US11035002B2 (en) 2012-08-14 2020-08-20 Methods and systems for processing polynucleotides
US16/998,414 US11021749B2 (en) 2012-08-14 2020-08-20 Methods and systems for processing polynucleotides
AU2020244615A AU2020244615B2 (en) 2013-06-27 2020-10-05 Compositions and methods for sample processing
US17/314,526 US11359239B2 (en) 2012-08-14 2021-05-07 Methods and systems for processing polynucleotides
US17/392,610 US20220098659A1 (en) 2012-08-14 2021-08-03 Methods and systems for processing polynucleotides
AU2022200718A AU2022200718B2 (en) 2012-08-14 2022-02-03 Microcapsule compositions and methods
US18/186,088 US20240002929A1 (en) 2012-08-14 2023-03-17 Methods and systems for processing polynucleotides
AU2023203879A AU2023203879A1 (en) 2013-06-27 2023-06-20 Compositions and methods for sample processing
US18/384,527 US20240376519A1 (en) 2012-08-14 2023-10-27 Compositions and methods for sample processing
AU2024203235A AU2024203235A1 (en) 2012-08-14 2024-05-15 Microcapsule compositions and methods
US18/760,903 US20250027149A1 (en) 2012-08-14 2024-07-01 Methods and systems for processing polynucleotides
US18/945,323 US20250066851A1 (en) 2012-08-14 2024-11-12 Methods and systems for processing polynucleotides
US18/960,947 US20250146047A1 (en) 2012-08-14 2024-11-26 Compositions and methods for sample processing

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US201261683192P 2012-08-14 2012-08-14
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US201361762435P 2013-02-08 2013-02-08
US61/762,435 2013-02-08
US201361800223P 2013-03-15 2013-03-15
US61/800,223 2013-03-15
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Cited By (127)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015157567A1 (en) 2014-04-10 2015-10-15 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
WO2015200871A1 (en) * 2014-06-26 2015-12-30 10X Genomics, Inc. Methods and compositions for sample analysis
US9290809B2 (en) 2009-12-15 2016-03-22 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US9315857B2 (en) 2009-12-15 2016-04-19 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse label-tags
US9388465B2 (en) 2013-02-08 2016-07-12 10X Genomics, Inc. Polynucleotide barcode generation
US9410201B2 (en) 2012-12-14 2016-08-09 10X Genomics, Inc. Methods and systems for processing polynucleotides
US20160244809A1 (en) * 2015-02-24 2016-08-25 10X Genomics, Inc. Partition Processing Methods and Systems
WO2016149547A1 (en) 2015-03-17 2016-09-22 Bio-Rad Laboratories, Inc. Detection of genome editing
WO2016172362A1 (en) * 2015-04-21 2016-10-27 General Automation Lab Technologies, Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
US9567645B2 (en) 2013-08-28 2017-02-14 Cellular Research, Inc. Massively parallel single cell analysis
KR20170020704A (ko) * 2014-06-26 2017-02-23 10엑스 제노믹스, 인크. 개별 세포 또는 세포 개체군으로부터 핵산을 분석하는 방법
US9582877B2 (en) 2013-10-07 2017-02-28 Cellular Research, Inc. Methods and systems for digitally counting features on arrays
US9689024B2 (en) 2012-08-14 2017-06-27 10X Genomics, Inc. Methods for droplet-based sample preparation
US9701998B2 (en) 2012-12-14 2017-07-11 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9727810B2 (en) 2015-02-27 2017-08-08 Cellular Research, Inc. Spatially addressable molecular barcoding
JP2017522049A (ja) * 2014-07-02 2017-08-10 ライフ テクノロジーズ コーポレーション センサー基質を装填するための方法
JP2017522866A (ja) * 2014-06-26 2017-08-17 １０エックス ジェノミクス， インコーポレイテッド 核酸配列の分析
US9824068B2 (en) 2013-12-16 2017-11-21 10X Genomics, Inc. Methods and apparatus for sorting data
WO2018027048A1 (en) 2016-08-05 2018-02-08 Bio-Rad Laboratories, Inc. Second strand direct
US9951386B2 (en) 2014-06-26 2018-04-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9975122B2 (en) 2014-11-05 2018-05-22 10X Genomics, Inc. Instrument systems for integrated sample processing
WO2018118971A1 (en) 2016-12-19 2018-06-28 Bio-Rad Laboratories, Inc. Droplet tagging contiguity preserved tagmented dna
US10011872B1 (en) 2016-12-22 2018-07-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
EP3013957B1 (en) 2013-06-27 2018-09-26 10X Genomics, Inc. Compositions and methods for sample processing
EP3271713A4 (en) * 2015-03-18 2018-10-24 The Broad Institute Inc. Massively parallel on-chip coalescence of microemulsions
US10202641B2 (en) 2016-05-31 2019-02-12 Cellular Research, Inc. Error correction in amplification of samples
US10221442B2 (en) 2012-08-14 2019-03-05 10X Genomics, Inc. Compositions and methods for sample processing
US10221436B2 (en) 2015-01-12 2019-03-05 10X Genomics, Inc. Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same
US10233487B2 (en) * 2016-03-24 2019-03-19 Bio-Rad Laboratories, Inc. Use of gel beads to control droplet dispersion
US10273541B2 (en) 2012-08-14 2019-04-30 10X Genomics, Inc. Methods and systems for processing polynucleotides
WO2019089959A1 (en) 2017-11-02 2019-05-09 Bio-Rad Laboratories, Inc. Transposase-based genomic analysis
US10287623B2 (en) 2014-10-29 2019-05-14 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequencing
US10301677B2 (en) 2016-05-25 2019-05-28 Cellular Research, Inc. Normalization of nucleic acid libraries
US10323279B2 (en) 2012-08-14 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
WO2019126789A1 (en) * 2017-12-22 2019-06-27 10X Genomics, Inc. Systems and methods for processing nucleic acid molecules from one or more cells
US10338066B2 (en) 2016-09-26 2019-07-02 Cellular Research, Inc. Measurement of protein expression using reagents with barcoded oligonucleotide sequences
EP3505933A1 (en) * 2017-12-29 2019-07-03 Blink AG A microcapsule for detecting and/or quantitating an analyte in a sample
WO2019148042A1 (en) * 2018-01-26 2019-08-01 10X Genomics, Inc. Compositions and methods for sample processing
WO2019152395A1 (en) 2018-01-31 2019-08-08 Bio-Rad Laboratories, Inc. Methods and compositions for deconvoluting partition barcodes
US10395758B2 (en) 2013-08-30 2019-08-27 10X Genomics, Inc. Sequencing methods
US10400235B2 (en) 2017-05-26 2019-09-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US10400280B2 (en) 2012-08-14 2019-09-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10428326B2 (en) 2017-01-30 2019-10-01 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
US10434507B2 (en) 2014-10-22 2019-10-08 The Regents Of The University Of California High definition microdroplet printer
US10501739B2 (en) 2017-10-18 2019-12-10 Mission Bio, Inc. Method, systems and apparatus for single cell analysis
US10518241B2 (en) * 2014-04-15 2019-12-31 Agilent Technologies, Inc. Creating and harvesting surface-bound emulsion
US10533221B2 (en) 2012-12-14 2020-01-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10550429B2 (en) 2016-12-22 2020-02-04 10X Genomics, Inc. Methods and systems for processing polynucleotides
WO2020041293A1 (en) 2018-08-20 2020-02-27 Bio-Rad Laboratories, Inc. Nucleotide sequence generation by barcode bead-colocalization in partitions
US10596541B2 (en) 2014-04-21 2020-03-24 President And Fellows Of Harvard College Systems and methods for barcoding nucleic acids
US10619186B2 (en) 2015-09-11 2020-04-14 Cellular Research, Inc. Methods and compositions for library normalization
US10640763B2 (en) 2016-05-31 2020-05-05 Cellular Research, Inc. Molecular indexing of internal sequences
US10650912B2 (en) 2015-01-13 2020-05-12 10X Genomics, Inc. Systems and methods for visualizing structural variation and phasing information
US10669570B2 (en) 2017-06-05 2020-06-02 Becton, Dickinson And Company Sample indexing for single cells
US10688494B2 (en) 2016-08-23 2020-06-23 10X Genomics, Inc. Microfluidic surface-mediated emulsion stability control
US10697010B2 (en) 2015-02-19 2020-06-30 Becton, Dickinson And Company High-throughput single-cell analysis combining proteomic and genomic information
US10697007B2 (en) 2014-06-27 2020-06-30 The Regents Of The University Of California PCR-activated sorting (PAS)
US10722880B2 (en) 2017-01-13 2020-07-28 Cellular Research, Inc. Hydrophilic coating of fluidic channels
US10745762B2 (en) 2012-08-13 2020-08-18 The Regents Of The University Of California Method and system for synthesizing a target polynucleotide within a droplet
US10745742B2 (en) 2017-11-15 2020-08-18 10X Genomics, Inc. Functionalized gel beads
US10752949B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10774370B2 (en) 2015-12-04 2020-09-15 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
US10815525B2 (en) 2016-12-22 2020-10-27 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10822643B2 (en) 2016-05-02 2020-11-03 Cellular Research, Inc. Accurate molecular barcoding
US10829815B2 (en) 2017-11-17 2020-11-10 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
US10839939B2 (en) 2014-06-26 2020-11-17 10X Genomics, Inc. Processes and systems for nucleic acid sequence assembly
US10854315B2 (en) 2015-02-09 2020-12-01 10X Genomics, Inc. Systems and methods for determining structural variation and phasing using variant call data
US10941396B2 (en) 2012-02-27 2021-03-09 Becton, Dickinson And Company Compositions and kits for molecular counting
US10947587B2 (en) * 2013-11-05 2021-03-16 The Regents Of The University Of California Single-cell forensic short tandem repeat typing within microfluidic droplets
US11001883B2 (en) 2012-03-05 2021-05-11 The General Hospital Corporation Systems and methods for epigenetic sequencing
WO2021146184A1 (en) * 2020-01-13 2021-07-22 Fluent Biosciences Inc. Single cell sequencing
US11081208B2 (en) 2016-02-11 2021-08-03 10X Genomics, Inc. Systems, methods, and media for de novo assembly of whole genome sequence data
US11084036B2 (en) 2016-05-13 2021-08-10 10X Genomics, Inc. Microfluidic systems and methods of use
EP3862090A1 (en) * 2020-02-10 2021-08-11 Roche Diagnostics GmbH Liquid dispensing system for a microfluidic sample carrier, microfluidic sample carrier sealing system including such liquid dispensing system, and method for dispensing sealing liquid using the same
US11111519B2 (en) 2015-02-04 2021-09-07 The Regents Of The University Of California Sequencing of nucleic acids via barcoding in discrete entities
US11124823B2 (en) 2015-06-01 2021-09-21 Becton, Dickinson And Company Methods for RNA quantification
US11124830B2 (en) 2016-12-21 2021-09-21 The Regents Of The University Of California Single cell genomic sequencing using hydrogel based droplets
US11142791B2 (en) 2016-08-10 2021-10-12 The Regents Of The University Of California Combined multiple-displacement amplification and PCR in an emulsion microdroplet
US11155881B2 (en) 2018-04-06 2021-10-26 10X Genomics, Inc. Systems and methods for quality control in single cell processing
US11155809B2 (en) 2014-06-24 2021-10-26 Bio-Rad Laboratories, Inc. Digital PCR barcoding
US11164659B2 (en) 2016-11-08 2021-11-02 Becton, Dickinson And Company Methods for expression profile classification
US11177020B2 (en) 2012-02-27 2021-11-16 The University Of North Carolina At Chapel Hill Methods and uses for molecular tags
CN113736850A (zh) * 2021-08-13 2021-12-03 纳昂达(南京)生物科技有限公司 基于双链环化的文库构建方法及其在测序中的应用
US11274343B2 (en) 2015-02-24 2022-03-15 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequence coverage
US11319583B2 (en) 2017-02-01 2022-05-03 Becton, Dickinson And Company Selective amplification using blocking oligonucleotides
US11331417B2 (en) 2015-09-29 2022-05-17 Shanghai Clinical Engine Technology Development Co., Ltd. Magnetic target separation instrument and application thereof
US11365409B2 (en) 2018-05-03 2022-06-21 Becton, Dickinson And Company Molecular barcoding on opposite transcript ends
US11365441B2 (en) 2019-05-22 2022-06-21 Mission Bio, Inc. Method and apparatus for simultaneous targeted sequencing of DNA, RNA and protein
US11371076B2 (en) 2019-01-16 2022-06-28 Becton, Dickinson And Company Polymerase chain reaction normalization through primer titration
US11390914B2 (en) 2015-04-23 2022-07-19 Becton, Dickinson And Company Methods and compositions for whole transcriptome amplification
US11397882B2 (en) 2016-05-26 2022-07-26 Becton, Dickinson And Company Molecular label counting adjustment methods
EP3797279A4 (en) * 2018-07-11 2022-08-03 The University of Hong Kong Automatic microfluidic system for continuous and quantitive collection of droplets
US11492660B2 (en) 2018-12-13 2022-11-08 Becton, Dickinson And Company Selective extension in single cell whole transcriptome analysis
US11512337B2 (en) 2020-01-13 2022-11-29 Fluent Biosciences Inc. Emulsion based drug screening
US11535882B2 (en) 2015-03-30 2022-12-27 Becton, Dickinson And Company Methods and compositions for combinatorial barcoding
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
US11608497B2 (en) 2016-11-08 2023-03-21 Becton, Dickinson And Company Methods for cell label classification
US11639517B2 (en) 2018-10-01 2023-05-02 Becton, Dickinson And Company Determining 5′ transcript sequences
US11649497B2 (en) 2020-01-13 2023-05-16 Becton, Dickinson And Company Methods and compositions for quantitation of proteins and RNA
US11661631B2 (en) 2019-01-23 2023-05-30 Becton, Dickinson And Company Oligonucleotides associated with antibodies
US11661625B2 (en) 2020-05-14 2023-05-30 Becton, Dickinson And Company Primers for immune repertoire profiling
US11667954B2 (en) 2019-07-01 2023-06-06 Mission Bio, Inc. Method and apparatus to normalize quantitative readouts in single-cell experiments
US11739443B2 (en) 2020-11-20 2023-08-29 Becton, Dickinson And Company Profiling of highly expressed and lowly expressed proteins
US11746367B2 (en) 2015-04-17 2023-09-05 President And Fellows Of Harvard College Barcoding systems and methods for gene sequencing and other applications
US11773436B2 (en) 2019-11-08 2023-10-03 Becton, Dickinson And Company Using random priming to obtain full-length V(D)J information for immune repertoire sequencing
US11773441B2 (en) 2018-05-03 2023-10-03 Becton, Dickinson And Company High throughput multiomics sample analysis
US11773389B2 (en) 2017-05-26 2023-10-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US11827936B2 (en) 2020-01-13 2023-11-28 Fluent Biosciences Inc. Methods and systems for single cell gene profiling
US11866782B2 (en) 2020-03-16 2024-01-09 Fluent Biosciences Inc. Multi-omic analysis in monodisperse droplets
US11898206B2 (en) 2017-05-19 2024-02-13 10X Genomics, Inc. Systems and methods for clonotype screening
US11932849B2 (en) 2018-11-08 2024-03-19 Becton, Dickinson And Company Whole transcriptome analysis of single cells using random priming
US11932901B2 (en) 2020-07-13 2024-03-19 Becton, Dickinson And Company Target enrichment using nucleic acid probes for scRNAseq
US11939622B2 (en) 2019-07-22 2024-03-26 Becton, Dickinson And Company Single cell chromatin immunoprecipitation sequencing assay
US11946095B2 (en) 2017-12-19 2024-04-02 Becton, Dickinson And Company Particles associated with oligonucleotides
US11965208B2 (en) 2019-04-19 2024-04-23 Becton, Dickinson And Company Methods of associating phenotypical data and single cell sequencing data
US12006543B2 (en) 2013-09-13 2024-06-11 Life Technologies Corporation Device preparation using condensed nucleic acid particles
US12071617B2 (en) 2019-02-14 2024-08-27 Becton, Dickinson And Company Hybrid targeted and whole transcriptome amplification
US12123888B2 (en) 2015-04-21 2024-10-22 Isolation Bio Inc. Apparatus and method for picking biological sample
US12153043B2 (en) 2020-02-25 2024-11-26 Becton, Dickinson And Company Bi-specific probes to enable the use of single-cell samples as single color compensation control
US12157913B2 (en) 2020-06-02 2024-12-03 Becton, Dickinson And Company Oligonucleotides and beads for 5 prime gene expression assay
US12188010B2 (en) 2020-01-29 2025-01-07 Becton, Dickinson And Company Barcoded wells for spatial mapping of single cells through sequencing
US12241059B2 (en) 2020-07-15 2025-03-04 Illumina, Inc. Tiered ligation oligos
US12264411B2 (en) 2017-01-30 2025-04-01 10X Genomics, Inc. Methods and systems for analysis
US12312640B2 (en) 2014-06-26 2025-05-27 10X Genomics, Inc. Analysis of nucleic acid sequences
US12392771B2 (en) 2020-12-15 2025-08-19 Becton, Dickinson And Company Single cell secretome analysis
US12391940B2 (en) 2020-07-31 2025-08-19 Becton, Dickinson And Company Single cell assay for transposase-accessible chromatin
US12421558B2 (en) 2021-02-12 2025-09-23 10X Genomics, Inc. Systems and methods for joint interactive visualization of gene expression and DNA chromatin accessibility

Families Citing this family (260)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9029085B2 (en) 2007-03-07 2015-05-12 President And Fellows Of Harvard College Assays and other reactions involving droplets
EP2235210B1 (en) 2007-12-21 2015-03-25 President and Fellows of Harvard College Methods for nucleic acid sequencing
WO2010033200A2 (en) 2008-09-19 2010-03-25 President And Fellows Of Harvard College Creation of libraries of droplets and related species
EP3587594B1 (en) 2008-12-19 2022-04-13 President and Fellows of Harvard College Particle-assisted nucleic acid sequencing
WO2010104597A2 (en) 2009-03-13 2010-09-16 President And Fellows Of Harvard College Scale-up of microfluidic devices
US10688026B2 (en) 2009-04-27 2020-06-23 Premier Dental Products Company Buffered microencapsulated compositions and methods
US9814657B2 (en) 2009-04-27 2017-11-14 Premier Dental Products Company Buffered microencapsulated compositions and methods
KR20120089661A (ko) 2009-09-02 2012-08-13 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 분출 및 다른 기술을 사용하여 생성되는 다중 에멀션
WO2011056546A1 (en) 2009-10-27 2011-05-12 President And Fellows Of Harvard College Droplet creation techniques
US10787701B2 (en) 2010-04-05 2020-09-29 Prognosys Biosciences, Inc. Spatially encoded biological assays
US20190300945A1 (en) 2010-04-05 2019-10-03 Prognosys Biosciences, Inc. Spatially Encoded Biological Assays
GB201106254D0 (en) 2011-04-13 2011-05-25 Frisen Jonas Method and product
EP3789498B1 (en) 2011-04-25 2025-08-13 Bio-Rad Laboratories, Inc. Methods for nucleic acid analysis
KR20140034242A (ko) 2011-05-23 2014-03-19 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 다중 에멀젼을 포함하는 에멀젼의 제어
ES2797448T3 (es) 2011-08-01 2020-12-02 Bio Rad Laboratories Sistema de captura de células
US10466160B2 (en) 2011-08-01 2019-11-05 Celsee Diagnostics, Inc. System and method for retrieving and analyzing particles
US9404864B2 (en) 2013-03-13 2016-08-02 Denovo Sciences, Inc. System for imaging captured cells
USRE50065E1 (en) 2012-10-17 2024-07-30 10X Genomics Sweden Ab Methods and product for optimising localised or spatial detection of gene expression in a tissue sample
US9752181B2 (en) 2013-01-26 2017-09-05 Denovo Sciences, Inc. System and method for capturing and analyzing cells
US9707562B2 (en) 2013-03-13 2017-07-18 Denovo Sciences, Inc. System for capturing and analyzing cells
US10391490B2 (en) 2013-05-31 2019-08-27 Celsee Diagnostics, Inc. System and method for isolating and analyzing cells
US9856535B2 (en) 2013-05-31 2018-01-02 Denovo Sciences, Inc. System for isolating cells
WO2014210225A1 (en) 2013-06-25 2014-12-31 Prognosys Biosciences, Inc. Methods and systems for determining spatial patterns of biological targets in a sample
US20160279068A1 (en) 2013-11-08 2016-09-29 President And Fellows Of Harvard College Microparticles, methods for their preparation and use
WO2015161177A1 (en) * 2014-04-17 2015-10-22 President And Fellows Of Harvard College Systems and methods for droplet tagging
JP2017532024A (ja) 2014-09-09 2017-11-02 ザ・ブロード・インスティテュート・インコーポレイテッド コンポジット単一細胞核酸分析のための液滴ベースの方法および機器
FR3031914B1 (fr) * 2015-01-27 2019-06-07 Calyxia Procede d'encapsulation
CN107614700A (zh) 2015-03-11 2018-01-19 布罗德研究所有限公司 基因型和表型偶联
ES2994053T3 (en) 2015-04-10 2025-01-17 10X Genomics Sweden Ab Spatially distinguished, multiplex nucleic acid analysis of biological specimens
EP4190912A1 (en) 2015-05-11 2023-06-07 Illumina, Inc. Platform for discovery and analysis of therapeutic agents
US9771575B2 (en) 2015-06-19 2017-09-26 Agilent Technologies, Inc. Methods for on-array fragmentation and barcoding of DNA samples
WO2017015099A1 (en) 2015-07-17 2017-01-26 Nanostring Technologies, Inc. Simultaneous quantification of gene expression in a user-defined region of a cross-sectioned tissue
US10974959B1 (en) * 2015-09-01 2021-04-13 Hrl Laboratories, Llc Methods for charge-titrating assembly of partially metallized nanoparticles, and metamaterials produced therefrom
US10189718B1 (en) * 2015-09-01 2019-01-29 Hrl Laboratories, Llc Methods for charge-titrating particle assembly, and structures produced therefrom
EP3362032A4 (en) 2015-10-13 2019-05-01 President and Fellows of Harvard College SYSTEMS AND METHOD FOR THE PRODUCTION AND USE OF GEL MICROBALLETS
EP3365454A1 (en) 2015-10-20 2018-08-29 10X Genomics, Inc. Methods and systems for high throughput single cell genetic manipulation
WO2017075297A1 (en) 2015-10-28 2017-05-04 The Broad Institute Inc. High-throughput dynamic reagent delivery system
US11092607B2 (en) 2015-10-28 2021-08-17 The Board Institute, Inc. Multiplex analysis of single cell constituents
DE102015221850A1 (de) * 2015-11-06 2017-05-11 Carl Zeiss Ag Verfahren zur Präparation von Referenzmarkierungen auf einem Probenträger
CN106755292B (zh) * 2015-11-19 2019-06-18 赛纳生物科技(北京)有限公司 一种磷酸修饰荧光团的核苷酸分子测序方法
US11371094B2 (en) 2015-11-19 2022-06-28 10X Genomics, Inc. Systems and methods for nucleic acid processing using degenerate nucleotides
EP3402902B1 (en) 2016-01-15 2021-10-27 Massachusetts Institute Of Technology Semi-permeable arrays for analyzing biological systems and methods of using same
SG11201809649WA (en) 2016-05-02 2018-11-29 Encodia Inc Macromolecule analysis employing nucleic acid encoding
JP2017203665A (ja) * 2016-05-10 2017-11-16 国立大学法人信州大学 光開裂性マイクロカプセル、それを用いたセンサ、及びそれを用いた被測定物質の測定方法
US11219919B1 (en) 2016-08-19 2022-01-11 Hrl Laboratories, Llc Electrostatically driven assembly of nanoparticle materials into dense films
CN109983126A (zh) 2016-10-19 2019-07-05 10X基因组学有限公司 用于条形码化单个细胞或细胞群的核酸分子的方法和系统
IL266197B2 (en) 2016-10-24 2024-03-01 Geneinfosec Inc Hiding information contained within nucleic acids
US10995333B2 (en) 2017-02-06 2021-05-04 10X Genomics, Inc. Systems and methods for nucleic acid preparation
EP3585364B1 (en) 2017-02-24 2021-04-28 The Regents of the University of California Particle-drop structures and methods for making and using the same
JP2020515871A (ja) 2017-03-21 2020-05-28 ミュウェルス，インコーポレイテッド 密封型マイクロウェルアッセイ
JP6911466B2 (ja) * 2017-03-31 2021-07-28 ソニーグループ株式会社 細胞の代謝を測定するためのマイクロチップ、細胞の代謝を測定するための装置、細胞の代謝を測定する方法及び細胞の代謝を測定するためのシステム
WO2018200867A1 (en) 2017-04-26 2018-11-01 10X Genomics, Inc. Mmlv reverse transcriptase variants
US11072816B2 (en) 2017-05-03 2021-07-27 The Broad Institute, Inc. Single-cell proteomic assay using aptamers
KR20200004335A (ko) * 2017-05-05 2020-01-13 스키피오 바이오사이언스 하이드로겔 내의 별개의 생물학적 단위를 트래핑하고 바코딩하는 방법
EP3403724A1 (en) * 2017-05-18 2018-11-21 Hifibio Process for manufacturing an array with microchannels
US10544413B2 (en) 2017-05-18 2020-01-28 10X Genomics, Inc. Methods and systems for sorting droplets and beads
EP4215616B1 (en) 2017-05-18 2024-09-04 10X Genomics, Inc. Methods and systems for sorting droplets and beads
WO2018226546A1 (en) 2017-06-05 2018-12-13 10X Genomics, Inc. Gaskets for the distribution of pressures in a microfluidic system
CN110799679A (zh) 2017-06-20 2020-02-14 10X基因组学有限公司 用于改善液滴稳定的方法和系统
US10498001B2 (en) 2017-08-21 2019-12-03 Texas Instruments Incorporated Launch structures for a hermetically sealed cavity
US10821442B2 (en) 2017-08-22 2020-11-03 10X Genomics, Inc. Devices, systems, and kits for forming droplets
AU2018323449B2 (en) 2017-08-29 2020-09-03 Bio-Rad Laboratories, Inc. System and method for isolating and analyzing cells
US10748643B2 (en) 2017-08-31 2020-08-18 10X Genomics, Inc. Systems and methods for determining the integrity of test strings with respect to a ground truth string
US10775422B2 (en) 2017-09-05 2020-09-15 Texas Instruments Incorporated Molecular spectroscopy cell with resonant cavity
US10589986B2 (en) 2017-09-06 2020-03-17 Texas Instruments Incorporated Packaging a sealed cavity in an electronic device
US10131115B1 (en) 2017-09-07 2018-11-20 Texas Instruments Incorporated Hermetically sealed molecular spectroscopy cell with dual wafer bonding
US10551265B2 (en) 2017-09-07 2020-02-04 Texas Instruments Incorporated Pressure sensing using quantum molecular rotational state transitions
US10424523B2 (en) 2017-09-07 2019-09-24 Texas Instruments Incorporated Hermetically sealed molecular spectroscopy cell with buried ground plane
US10549986B2 (en) 2017-09-07 2020-02-04 Texas Instruments Incorporated Hermetically sealed molecular spectroscopy cell
US10444102B2 (en) 2017-09-07 2019-10-15 Texas Instruments Incorporated Pressure measurement based on electromagnetic signal output of a cavity
US10544039B2 (en) * 2017-09-08 2020-01-28 Texas Instruments Incorporated Methods for depositing a measured amount of a species in a sealed cavity
EP3691703A1 (en) 2017-10-04 2020-08-12 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US10837047B2 (en) 2017-10-04 2020-11-17 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US10590244B2 (en) 2017-10-04 2020-03-17 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
WO2019084043A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS AND SYSTEMS FOR NUCLEIC ACID PREPARATION AND CHROMATIN ANALYSIS
WO2019084328A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS FOR PREPARING NUCLEIC ACID MOLECULES
WO2019083852A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. MICROFLUIDIC CHANNEL NETWORKS FOR PARTITIONING
EP4241882B1 (en) 2017-10-27 2025-04-30 10X Genomics, Inc. Methods for sample preparation and analysis
US11513126B2 (en) 2017-10-31 2022-11-29 Encodia, Inc. Kits for analysis using nucleic acid encoding and/or label
EP3559272B1 (en) 2017-11-17 2022-08-31 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
WO2019108851A1 (en) 2017-11-30 2019-06-06 10X Genomics, Inc. Systems and methods for nucleic acid preparation and analysis
WO2019113235A1 (en) 2017-12-06 2019-06-13 10X Genomics, Inc. Methods and systems for processing nucleic acid molecules
EP4403644A3 (en) 2017-12-08 2024-10-16 10X Genomics, Inc. Methods and compositions for labeling cells
CN111699388B (zh) 2017-12-12 2024-08-02 10X基因组学有限公司 用于单细胞处理的系统和方法
WO2019126466A1 (en) * 2017-12-22 2019-06-27 Rootpath Genomics, Inc. Compositions and methods for barcoding
CN112004920B (zh) 2018-02-05 2024-08-02 斯坦福大学托管董事会 用于单细胞和集合细胞的多重测量的系统和方法
CA3090699A1 (en) 2018-02-12 2019-08-15 Nanostring Technologies, Inc. Biomolecular probes and methods of detecting gene and protein expression
SG11202007686VA (en) 2018-02-12 2020-09-29 10X Genomics Inc Methods characterizing multiple analytes from individual cells or cell populations
EP3755812A1 (en) 2018-02-22 2020-12-30 10X Genomics, Inc. Ligation mediated analysis of nucleic acids
US11639928B2 (en) 2018-02-22 2023-05-02 10X Genomics, Inc. Methods and systems for characterizing analytes from individual cells or cell populations
WO2019169028A1 (en) 2018-02-28 2019-09-06 10X Genomics, Inc. Transcriptome sequencing through random ligation
CA3093958A1 (en) * 2018-03-14 2019-09-19 Premier Dental Products Company Buffered microencapsulated compositions and methods
WO2019191321A1 (en) 2018-03-28 2019-10-03 10X Genomics, Inc. Nucleic acid enrichment within partitions
WO2019209290A1 (en) 2018-04-25 2019-10-31 Hewlett-Packard Development Company, L.P. Selective release of material in thermally degradable capsule
WO2019209308A1 (en) * 2018-04-27 2019-10-31 Hewlett-Packard Development Company, L.P. Thermally controlled reagent release
WO2019212536A1 (en) 2018-05-01 2019-11-07 Hewlett-Packard Development Company, L.P. Sequential encapsulation of reagents
WO2019217758A1 (en) 2018-05-10 2019-11-14 10X Genomics, Inc. Methods and systems for molecular library generation
WO2019219757A1 (de) * 2018-05-15 2019-11-21 Albert-Ludwig-Universität Freiburg Mikroarray-transformer
EP3797171B1 (en) * 2018-05-21 2025-08-27 The Regents of The University of California Single cell mapping and transcriptome analysis
US11932899B2 (en) 2018-06-07 2024-03-19 10X Genomics, Inc. Methods and systems for characterizing nucleic acid molecules
US11703427B2 (en) 2018-06-25 2023-07-18 10X Genomics, Inc. Methods and systems for cell and bead processing
US12188014B1 (en) 2018-07-25 2025-01-07 10X Genomics, Inc. Compositions and methods for nucleic acid processing using blocking agents
US20200032335A1 (en) 2018-07-27 2020-01-30 10X Genomics, Inc. Systems and methods for metabolome analysis
SG11202101164TA (en) 2018-08-03 2021-03-30 10X Genomics Inc Methods and systems to minimize barcode exchange
IL280682B (en) 2018-08-13 2022-08-01 Rootpath Genomics Inc High throughput cloning of paired bipartite immunoreceptor polynucleotides and applications thereof
US20210268465A1 (en) * 2018-08-17 2021-09-02 The Regents Of The University Of California Particle-containing droplet systems with monodisperse fluid volumes
US12065688B2 (en) 2018-08-20 2024-08-20 10X Genomics, Inc. Compositions and methods for cellular processing
WO2020041148A1 (en) 2018-08-20 2020-02-27 10X Genomics, Inc. Methods and systems for detection of protein-dna interactions using proximity ligation
WO2020047005A1 (en) 2018-08-28 2020-03-05 10X Genomics, Inc. Resolving spatial arrays
EP3844304B1 (en) 2018-08-28 2024-10-02 10X Genomics, Inc. Methods for generating spatially barcoded arrays
US11519033B2 (en) 2018-08-28 2022-12-06 10X Genomics, Inc. Method for transposase-mediated spatial tagging and analyzing genomic DNA in a biological sample
FR3086951B1 (fr) * 2018-10-05 2021-02-26 Redberry Methode et dispositif pour la detection d'au moins un microorganisme selon sa cinetique de marquage, et support de detection
US11459607B1 (en) 2018-12-10 2022-10-04 10X Genomics, Inc. Systems and methods for processing-nucleic acid molecules from a single cell using sequential co-partitioning and composite barcodes
EP3894587A1 (en) 2018-12-10 2021-10-20 10X Genomics, Inc. Resolving spatial arrays by proximity-based deconvolution
WO2020123657A2 (en) 2018-12-11 2020-06-18 10X Genomics, Inc. Methods and devices for detecting and sorting droplets or particles
DE102019130236A1 (de) * 2018-12-20 2020-06-25 Endress+Hauser Conducta Gmbh+Co. Kg Verfahren und System zum Herstellen einer Lösung
WO2020139844A1 (en) 2018-12-24 2020-07-02 10X Genomics, Inc. Devices, systems, and methods for controlling liquid flow
TWI725686B (zh) 2018-12-26 2021-04-21 財團法人工業技術研究院 用於產生液珠的管狀結構及液珠產生方法
US11649485B2 (en) 2019-01-06 2023-05-16 10X Genomics, Inc. Generating capture probes for spatial analysis
EP3906318A1 (en) 2019-01-06 2021-11-10 10X Genomics, Inc. Methods and systems for enrichment of barcodes
US11926867B2 (en) 2019-01-06 2024-03-12 10X Genomics, Inc. Generating capture probes for spatial analysis
US12169198B2 (en) 2019-01-08 2024-12-17 10X Genomics, Inc. Systems and methods for sample analysis
US11845983B1 (en) 2019-01-09 2023-12-19 10X Genomics, Inc. Methods and systems for multiplexing of droplet based assays
US20200232979A1 (en) * 2019-01-22 2020-07-23 Mayo Foundation For Medical Education And Research Microcapsules and Methods for Analyte Detection
JP7528103B2 (ja) * 2019-02-11 2024-08-05 ウルティマ・ゲノミクス・インコーポレーテッド 核酸分析方法
JP7448974B2 (ja) 2019-02-12 2024-03-13 セリーセル ゲーエムベーハー 逆免疫抑制
US11467153B2 (en) 2019-02-12 2022-10-11 10X Genomics, Inc. Methods for processing nucleic acid molecules
US12305239B2 (en) 2019-02-12 2025-05-20 10X Genomics, Inc. Analysis of nucleic acid sequences
CN113614245B (zh) 2019-02-12 2024-08-27 10X基因组学有限公司 用于加工核酸分子的方法
WO2020167862A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transfer of reagents between droplets
US11851683B1 (en) 2019-02-12 2023-12-26 10X Genomics, Inc. Methods and systems for selective analysis of cellular samples
WO2020167866A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transposon loading
US12275993B2 (en) 2019-02-12 2025-04-15 10X Genomics, Inc. Analysis of nucleic acid sequences
US11655499B1 (en) 2019-02-25 2023-05-23 10X Genomics, Inc. Detection of sequence elements in nucleic acid molecules
WO2020176882A1 (en) 2019-02-28 2020-09-03 10X Genomics, Inc. Devices, systems, and methods for increasing droplet formation efficiency
WO2020176788A1 (en) 2019-02-28 2020-09-03 10X Genomics, Inc. Profiling of biological analytes with spatially barcoded oligonucleotide arrays
SG11202111242PA (en) 2019-03-11 2021-11-29 10X Genomics Inc Systems and methods for processing optically tagged beads
CN114127309A (zh) 2019-03-15 2022-03-01 10X基因组学有限公司 使用空间阵列进行单细胞测序的方法
EP3941630B1 (en) * 2019-03-18 2025-04-16 Cellular Research, Inc. Precise delivery of components into fluids
WO2020198071A1 (en) 2019-03-22 2020-10-01 10X Genomics, Inc. Three-dimensional spatial analysis
EP3947722A1 (en) 2019-03-27 2022-02-09 10X Genomics, Inc. Systems and methods for processing rna from cells
EP3947530A1 (en) 2019-04-03 2022-02-09 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
KR20220004060A (ko) 2019-04-05 2022-01-11 루트패스 제노믹스, 인크. T-세포 수용체 유전자 어셈블리를 위한 조성물 및 방법
WO2020207963A1 (en) 2019-04-12 2020-10-15 Miltenyi Biotec B.V. & Co. KG Conjugates having an enzymatically releasable detection moiety and a barcode moiety
US10633693B1 (en) * 2019-04-16 2020-04-28 Celsee Diagnostics, Inc. System and method for leakage control in a particle capture system
JPWO2020218551A1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) * 2019-04-26 2020-10-29
US11273439B2 (en) 2019-05-07 2022-03-15 Bio-Rad Laboratories, Inc. System and method for target material retrieval from microwells
CA3138881A1 (en) 2019-05-07 2020-11-12 Bio-Rad Laboratories, Inc. System and method for automated single cell processing
EP3976820A1 (en) 2019-05-30 2022-04-06 10X Genomics, Inc. Methods of detecting spatial heterogeneity of a biological sample
CN118291246A (zh) 2019-06-14 2024-07-05 伯乐实验室有限公司 用于自动化单细胞处理和分析的系统和方法
US12186751B2 (en) 2019-06-28 2025-01-07 10X Genomics, Inc. Devices and systems incorporating acoustic ordering and methods of use thereof
US11919002B2 (en) 2019-08-20 2024-03-05 10X Genomics, Inc. Devices and methods for generating and recovering droplets
CN114729392A (zh) 2019-09-06 2022-07-08 10X基因组学有限公司 用于对细胞和细胞珠粒进行条形码化的系统和方法
US12235262B1 (en) 2019-09-09 2025-02-25 10X Genomics, Inc. Methods and systems for single cell protein analysis
AU2020361681A1 (en) 2019-10-10 2022-05-05 1859, Inc. Methods and systems for microfluidic screening
CN114761800A (zh) 2019-10-11 2022-07-15 10X基因组学有限公司 用于分析物检测和分析的方法
US12157124B2 (en) 2019-11-06 2024-12-03 10X Genomics, Inc. Imaging system hardware
WO2021091611A1 (en) 2019-11-08 2021-05-14 10X Genomics, Inc. Spatially-tagged analyte capture agents for analyte multiplexing
EP4058596A1 (en) 2019-11-15 2022-09-21 Miltenyi Biotec B.V. & Co. KG Color and bardcoded beads for single cell indexing
US12059679B2 (en) 2019-11-19 2024-08-13 10X Genomics, Inc. Methods and devices for sorting droplets and particles
CN114901395B (zh) 2019-11-27 2024-11-05 10X基因组学有限公司 用于产生液滴的装置、系统和方法
WO2021119201A1 (en) 2019-12-10 2021-06-17 Enumerix, Inc. Methods and systems for three-dimensional lightsheet imaging
EP4073241A2 (en) 2019-12-11 2022-10-19 10X Genomics, Inc. Reverse transcriptase variants
US12241890B2 (en) 2019-12-23 2025-03-04 10X Genomics, Inc. Methods for generating barcoded nucleic acid molecules using fixed cells
US20210214769A1 (en) * 2020-01-13 2021-07-15 Fluent Biosciences Inc. Methods and systems for amplifying low concentrations of nucleic acids
US12405264B2 (en) 2020-01-17 2025-09-02 10X Genomics, Inc. Electrophoretic system and method for analyte capture
US11702693B2 (en) 2020-01-21 2023-07-18 10X Genomics, Inc. Methods for printing cells and generating arrays of barcoded cells
US11732299B2 (en) 2020-01-21 2023-08-22 10X Genomics, Inc. Spatial assays with perturbed cells
US20210230681A1 (en) 2020-01-24 2021-07-29 10X Genomics, Inc. Methods for spatial analysis using proximity ligation
US12112833B2 (en) 2020-02-04 2024-10-08 10X Genomics, Inc. Systems and methods for index hopping filtering
US11835462B2 (en) 2020-02-11 2023-12-05 10X Genomics, Inc. Methods and compositions for partitioning a biological sample
WO2021163611A1 (en) 2020-02-13 2021-08-19 10X Genomics, Inc. Methods for characterizing cells using gene expression and chromatin accessibility
US11926863B1 (en) 2020-02-27 2024-03-12 10X Genomics, Inc. Solid state single cell method for analyzing fixed biological cells
CN115461470A (zh) 2020-02-28 2022-12-09 10X基因组学有限公司 用于从组织中分离核和细胞的方法
US11504719B2 (en) 2020-03-12 2022-11-22 Bio-Rad Laboratories, Inc. System and method for receiving and delivering a fluid for sample processing
EP4121528A1 (en) 2020-03-17 2023-01-25 F. Hoffmann-La Roche AG Method of improving cell recovery in single-cell analysis
WO2021212042A1 (en) 2020-04-16 2021-10-21 10X Genomics, Inc. Compositions and methods for use with fixed samples
WO2021222302A1 (en) 2020-04-27 2021-11-04 10X Genomics, Inc. Methods and systems for increasing cell recovery efficiency
WO2021222301A1 (en) 2020-04-27 2021-11-04 10X Genomics, Inc. Methods and systems for analysis and identification of barcode multiplets
WO2021226290A1 (en) 2020-05-05 2021-11-11 10X Genomics, Inc. Methods for identification of antigen-binding molecules
US11851700B1 (en) 2020-05-13 2023-12-26 10X Genomics, Inc. Methods, kits, and compositions for processing extracellular molecules
CN116134308B (zh) 2020-05-19 2025-05-27 10X基因组学有限公司 电泳盒和仪器
ES2989052T3 (es) 2020-05-22 2024-11-25 10X Genomics Inc Medición espacio-temporal simultánea de la expresión génica y la actividad celular
EP4158058B1 (en) 2020-06-02 2025-08-06 10X Genomics, Inc. Enrichment of nucleic acid sequences
US12265079B1 (en) 2020-06-02 2025-04-01 10X Genomics, Inc. Systems and methods for detecting analytes from captured single biological particles
US12031177B1 (en) 2020-06-04 2024-07-09 10X Genomics, Inc. Methods of enhancing spatial resolution of transcripts
US12168801B1 (en) 2020-07-02 2024-12-17 10X Genomics, Inc. Hybrid/capture probe designs for full-length cDNA
US11434525B2 (en) 2020-08-06 2022-09-06 Singular Genomics Systems, Inc. Spatial sequencing
US11492662B2 (en) 2020-08-06 2022-11-08 Singular Genomics Systems, Inc. Methods for in situ transcriptomics and proteomics
WO2022051522A1 (en) 2020-09-02 2022-03-10 10X Genomics, Inc. Flow focusing devices, systems, and methods for high throughput droplet formation
CN116171200A (zh) 2020-09-02 2023-05-26 10X基因组学有限公司 用于高通量液滴形成的装置、系统和方法
JP7612841B2 (ja) 2020-09-07 2025-01-14 ミルテニー バイオテック ベー．フェー． ウント コー． カー・ゲー 酵素的に放出可能な検出部分およびバーコード部分を有するコンジュゲート
AU2021345283B2 (en) 2020-09-18 2024-12-19 10X Genomics, Inc. Sample handling apparatus and image registration methods
EP4410956A3 (en) 2020-09-23 2025-04-09 10X Genomics, Inc. Selective enzymatic gelation
US12298314B2 (en) 2020-10-09 2025-05-13 10X Genomics, Inc. Methods and compositions for analyzing target binding of molecules
CN116964219A (zh) 2020-10-09 2023-10-27 10X基因组学有限公司 用于分析抗原结合分子的方法和组合物
WO2022076914A1 (en) 2020-10-09 2022-04-14 10X Genomics, Inc. Methods and compositions for profiling immune repertoire
WO2022081643A2 (en) 2020-10-13 2022-04-21 10X Genomics, Inc. Compositions and methods for generating recombinant antigen binding molecules from single cells
WO2022096294A2 (en) 2020-11-03 2022-05-12 Droplet Genomics, Uab Integrated platform for selective microfluidic particle processing
US12084715B1 (en) 2020-11-05 2024-09-10 10X Genomics, Inc. Methods and systems for reducing artifactual antisense products
EP4001432A1 (en) 2020-11-13 2022-05-25 Miltenyi Biotec B.V. & Co. KG Algorithmic method for efficient indexing of genetic sequences using associative arrays
WO2022150662A1 (en) 2021-01-08 2022-07-14 10X Genomics, Inc. Methods for generating antigen-binding molecules from single cells
US12398262B1 (en) 2021-01-22 2025-08-26 10X Genomics, Inc. Triblock copolymer-based cell stabilization and fixation system and methods of use thereof
US12390775B1 (en) 2021-02-08 2025-08-19 10X Genomics, Inc. Devices and methods for reducing the effects of settling of particles during droplet production
WO2022178304A1 (en) 2021-02-19 2022-08-25 10X Genomics, Inc. High-throughput methods for analyzing and affinity-maturing an antigen-binding molecule
EP4421491A3 (en) 2021-02-19 2024-11-27 10X Genomics, Inc. Method of using a modular assay support device
WO2022182682A1 (en) 2021-02-23 2022-09-01 10X Genomics, Inc. Probe-based analysis of nucleic acids and proteins
EP4298239A1 (en) 2021-02-23 2024-01-03 10X Genomics, Inc. Drug screening methods
WO2022182664A1 (en) 2021-02-23 2022-09-01 10X Genomics, Inc. A method for epitope binning of novel monoclonal antibodies
WO2022182662A1 (en) 2021-02-23 2022-09-01 10X Genomics, Inc. Compositions and methods for mapping antigen-binding molecule affinity to antigen regions of interest
CN117178061A (zh) 2021-02-23 2023-12-05 10X基因组学有限公司 单细胞聚糖谱分析
EP4297901A1 (en) 2021-02-24 2024-01-03 10X Genomics, Inc. Method for concentrating droplets in an emulsion
EP4301499A4 (en) 2021-03-05 2025-01-22 Enumerix, Inc. SYSTEMS AND METHODS FOR GENERATING DROPLETS AND PERFORMING DIGITAL ANALYSES
WO2022204539A1 (en) 2021-03-26 2022-09-29 10X Genomics, Inc. Devices, methods, and systems for improved droplet recovery
WO2022221428A1 (en) 2021-04-14 2022-10-20 10X Genomics, Inc. Compositions and methods for single cell analyte detection and analysis
CN117440860A (zh) * 2021-04-16 2024-01-23 普莱克斯姆公司 用于组合文库的测定装置
WO2022220843A1 (en) * 2021-04-16 2022-10-20 Plexium, Inc. Caps for assay devices
EP4334706A4 (en) 2021-05-05 2025-08-06 Singular Genomics Systems Inc MULTIOMICS ANALYSIS DEVICE AND METHODS OF USE THEREOF
CA3220479A1 (en) * 2021-05-18 2022-11-24 Fluent Biosciences Inc. Template particles with micropores and nanopores
WO2022256313A1 (en) 2021-06-01 2022-12-08 10X Genomics, Inc. Validation of a unique molecular identifier associated with a nucleic acid sequence of interest
WO2022256345A1 (en) 2021-06-01 2022-12-08 10X Genomics, Inc. Methods and systems for engineering antibodies, and antigen-binding fragments thereof, to have altered characteristics
EP4373966B1 (en) 2021-07-26 2025-04-09 10X Genomics, Inc. Nucleic acid processing via circularization
WO2023022925A1 (en) 2021-08-17 2023-02-23 10X Genomics, Inc. Compositions, systems and methods for enzyme optimization
US12252745B2 (en) 2021-09-02 2025-03-18 Enumerix, Inc. Detection and digital quantitation of multiple targets
USD1064308S1 (en) 2021-09-17 2025-02-25 10X Genomics, Inc. Sample handling device
WO2023060110A1 (en) 2021-10-05 2023-04-13 10X Genomics, Inc. Methods of immune cell analysis
WO2023076832A1 (en) 2021-10-25 2023-05-04 Singular Genomics Systems, Inc. Manipulating and detecting biological samples
EP4448787A1 (en) 2021-12-13 2024-10-23 Cornell University Genotyping of targeted loci with single-cell chromatin accessibility
WO2023114310A1 (en) 2021-12-15 2023-06-22 10X Genomics, Inc. Methods for improving sensitivity of immune profiling using oligo-tagged antigens
WO2023122033A1 (en) 2021-12-20 2023-06-29 10X Genomics, Inc. Self-test for pathology/histology slide imaging device
EP4453246A1 (en) 2021-12-20 2024-10-30 Enumerix, Inc. Detection and digital quantitation of multiple targets
EP4486817A1 (en) 2022-03-04 2025-01-08 10x Genomics, Inc. Droplet forming devices and methods having fluoropolymer silane coating agents
WO2023215320A1 (en) * 2022-05-02 2023-11-09 The Brigham And Women’S Hospital, Inc. Microfluidic platform for the study of molecular and/or cellular interactions
WO2023215861A1 (en) 2022-05-06 2023-11-09 10X Genomics, Inc. Reagents for characterizing antigen-binding molecules from immune cells
WO2023225259A1 (en) 2022-05-20 2023-11-23 10X Genomics, Inc. Compositions and methods for characterizing antigen binding molecules from single cells
WO2023225201A1 (en) 2022-05-20 2023-11-23 10X Genomics, Inc. Compositions and methods for characterizing t cell, or t cell-like, receptors from single cells
WO2023225294A1 (en) 2022-05-20 2023-11-23 10X Genomics, Inc. Improved major histocompatibility complex molecules
EP4532747A1 (en) * 2022-06-01 2025-04-09 Bio-Rad Laboratories, Inc. Method and system for detecting reverse transcriptase activity by digital assay
WO2023235596A1 (en) 2022-06-03 2023-12-07 10X Genomics, Inc. Systems and methods for determining antigen binding specificity of antigen binding molecules
WO2023235570A1 (en) 2022-06-03 2023-12-07 10X Genomics, Inc. Methods and compositions for the identification of antigen binding molecules using lipoparticle-based antigen mapping
EP4536171A2 (en) * 2022-06-10 2025-04-16 Ohio State Innovation Foundation Sustained adjunct therapy to improve chemotherapy efficacy in glioblastoma in a cerebrovascular-tumor-on-a-chip model
WO2023250422A1 (en) 2022-06-23 2023-12-28 10X Genomics, Inc. Compositions and methods for characterizing multispecific antigen binding molecules from single cells
WO2024006734A1 (en) 2022-06-27 2024-01-04 10X Genomics, Inc. Methods for preparing and using mhc multimer reagents compositions
WO2024006392A1 (en) 2022-06-29 2024-01-04 10X Genomics, Inc. Probe-based analysis of nucleic acids and proteins
WO2024015856A1 (en) 2022-07-13 2024-01-18 10X Genomics, Inc. Compositions and methods for characterizing binding characteristics of antigen binding molecules from single cells
WO2024015378A1 (en) 2022-07-13 2024-01-18 10X Genomics, Inc. Methods and systems for characterizing antigen-binding molecules expressed by immune cells
WO2024015733A1 (en) 2022-07-13 2024-01-18 10X Genomics, Inc. Improved methods and systems for identification and characterization of antigen-binding molecules from single cells
CN118201982A (zh) 2022-08-18 2024-06-14 10X基因组学有限公司 具有含氟二醇添加剂的液滴形成装置和方法
WO2024050299A1 (en) 2022-08-29 2024-03-07 10X Genomics, Inc. Improved methods and compositions for characterization of antigen-binding molecules from single cells
US20250257399A1 (en) 2022-10-03 2025-08-14 10X Genomics, Inc. Compositions and methods for analyzing genomic insertion sites of exogenous nucleic acids
WO2024098046A1 (en) 2022-11-04 2024-05-10 10X Genomics, Inc. Systems and methods for determining antigen specificity of antigen binding molecules and visualizing adaptive immune cell clonotyping data
WO2024243444A1 (en) 2023-05-23 2024-11-28 10X Genomics, Inc. Methods and systems for characterizing analytes from individual cells or cell populations
WO2024249961A1 (en) 2023-06-01 2024-12-05 Singular Genomics Systems, Inc. Methods and probes for detecting polynucleotide sequences in cells and tissues
CN116905093B (zh) * 2023-06-07 2024-05-24 祥符实验室 一种dna数据库的长期保存方法及其应用
WO2025064703A1 (en) 2023-09-21 2025-03-27 10X Genomics, Inc. Compositions and methods for particle hashing
WO2025096581A1 (en) 2023-10-31 2025-05-08 10X Genomics, Inc. Methods of spatial analysis and single nuclei sequencing
WO2025111369A1 (en) 2023-11-21 2025-05-30 10X Genomics, Inc. Probe-based analysis of polynucleotides and proteins
WO2025166072A1 (en) 2024-02-01 2025-08-07 10X Genomics, Inc. Compositions and methods for sample analysis
WO2025166185A1 (en) 2024-02-02 2025-08-07 10X Genomics, Inc. Methods of crispr guide rna sequencing in single cell workflows
CN119184929A (zh) * 2024-10-10 2024-12-27 上海心玮医疗科技股份有限公司 一种快速治疗颅内血管狭窄的药物支架

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004010106A2 (en) * 2002-07-24 2004-01-29 Ptc Therapeutics, Inc. METHODS FOR IDENTIFYING SMALL MOLEDULES THAT MODULATE PREMATURE TRANSLATION TERMINATION AND NONSENSE MEDIATED mRNA DECAY
WO2006030993A1 (en) * 2004-09-14 2006-03-23 Jin-Ho Choy Information code system using dna sequences
US20090053169A1 (en) * 2007-08-20 2009-02-26 Pharmain Corporation Oligonucleotide Core Carrier Compositions for Delivery of Nucleic Acid-Containing Therapeutic Agents, Methods of Making and Using the Same
WO2010004018A2 (en) * 2008-07-11 2010-01-14 Eth Zurich Degradable microcapsules
WO2011066476A1 (en) * 2009-11-25 2011-06-03 Quantalife, Inc. Methods and compositions for detecting genetic material

Family Cites Families (823)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2797149A (en) 1953-01-08 1957-06-25 Technicon International Ltd Methods of and apparatus for analyzing liquids containing crystalloid and non-crystalloid constituents
US3047367A (en) 1959-12-01 1962-07-31 Technicon Instr Automatic analysis with fluid segmentation
US3479141A (en) 1967-05-17 1969-11-18 Technicon Corp Method and apparatus for analysis
US4124638A (en) 1977-09-12 1978-11-07 Hansen John N Solubilizable polyacrylamide gels containing disulfide cross-linkages
US4253846A (en) 1979-11-21 1981-03-03 Technicon Instruments Corporation Method and apparatus for automated analysis of fluid samples
GB2097692B (en) 1981-01-10 1985-05-22 Shaw Stewart P D Combining chemical reagents
DE3230289A1 (de) 1982-08-14 1984-02-16 Bayer Ag, 5090 Leverkusen Herstellung von pharmazeutischen oder kosmetischen dispersionen
US4582802A (en) 1983-09-30 1986-04-15 The United States Of America As Represented By The Department Of Health And Human Services Stimulation of enzymatic ligation of DNA by high concentrations of nonspecific polymers
JPS60227826A (ja) * 1984-04-27 1985-11-13 Sogo Yatsukou Kk ｐＨに応答するグラフトカプセル
KR890003947B1 (ko) 1985-12-11 1989-10-13 가부시기가이샤 시마즈세이사구쇼 세포융합장치
US4916070A (en) 1986-04-14 1990-04-10 The General Hospital Corporation Fibrin-specific antibodies and method of screening for the antibodies
US5618711A (en) 1986-08-22 1997-04-08 Hoffmann-La Roche Inc. Recombinant expression vectors and purification methods for Thermus thermophilus DNA polymerase
US4872895A (en) 1986-12-11 1989-10-10 American Telephone And Telegraph Company, At&T Bell Laboratories Method for fabricating articles which include high silica glass bodies
US5525464A (en) 1987-04-01 1996-06-11 Hyseq, Inc. Method of sequencing by hybridization of oligonucleotide probes
US5202231A (en) 1987-04-01 1993-04-13 Drmanac Radoje T Method of sequencing of genomes by hybridization of oligonucleotide probes
US5149625A (en) 1987-08-11 1992-09-22 President And Fellows Of Harvard College Multiplex analysis of DNA
US5137829A (en) 1987-10-05 1992-08-11 Washington University DNA transposon TN5SEQ1
US5185099A (en) 1988-04-20 1993-02-09 Institut National De Recherche Chimique Appliquee Visco-elastic, isotropic materials based on water, fluorinate sufactants and fluorinated oils, process for their preparation, and their use in various fields, such as optics, pharmacology and electrodynamics
US5237016A (en) 1989-01-05 1993-08-17 Siska Diagnostics, Inc. End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids
US6176962B1 (en) 1990-02-28 2001-01-23 Aclara Biosciences, Inc. Methods for fabricating enclosed microchannel structures
US5756334A (en) 1990-04-26 1998-05-26 New England Biolabs, Inc. Thermostable DNA polymerase from 9°N-7 and methods for producing the same
AU665338B2 (en) 1990-07-24 1996-01-04 F. Hoffmann-La Roche Ag The reduction of non-specific amplification during (in vitro) nucleic acid amplification using modified nucleic acid bases
US5489523A (en) 1990-12-03 1996-02-06 Stratagene Exonuclease-deficient thermostable Pyrococcus furiosus DNA polymerase I
US6582908B2 (en) 1990-12-06 2003-06-24 Affymetrix, Inc. Oligonucleotides
US5270183A (en) 1991-02-08 1993-12-14 Beckman Research Institute Of The City Of Hope Device and method for the automated cycling of solutions between two or more temperatures
US5994056A (en) 1991-05-02 1999-11-30 Roche Molecular Systems, Inc. Homogeneous methods for nucleic acid amplification and detection
WO1993001498A1 (en) 1991-07-04 1993-01-21 Immunodex K/S Water-soluble, polymer-based reagents and conjugates comprising moieties derived from divinyl sulfone
DE69228247T2 (de) * 1991-08-10 1999-07-08 Medical Research Council, London Behandlung von Zellpopulationen
WO1993006121A1 (en) 1991-09-18 1993-04-01 Affymax Technologies N.V. Method of synthesizing diverse collections of oligomers
US5413924A (en) 1992-02-13 1995-05-09 Kosak; Kenneth M. Preparation of wax beads containing a reagent for release by heating
WO1993019205A1 (en) 1992-03-19 1993-09-30 The Regents Of The University Of California Multiple tag labeling method for dna sequencing
US5587128A (en) 1992-05-01 1996-12-24 The Trustees Of The University Of Pennsylvania Mesoscale polynucleotide amplification devices
CA2134475C (en) 1992-05-01 1999-07-06 Peter Wilding Polynucleotide amplification analysis using a microfabricated device
US5840865A (en) 1992-09-14 1998-11-24 Institute Of Molecular Biology And Biotechnology/Forth Eukaryotic transposable element
US5897783A (en) 1992-09-24 1999-04-27 Amersham International Plc Magnetic separation method
US5569364A (en) 1992-11-05 1996-10-29 Soane Biosciences, Inc. Separation media for electrophoresis
AU673245B2 (en) 1993-02-01 1996-10-31 Seq, Ltd. Methods and apparatus for DNA sequencing
AU6175594A (en) 1993-02-16 1994-09-14 Alliance Pharmaceutical Corporation Method of microemulsifying fluorinated oils
EP0695170A4 (en) 1993-04-19 1998-08-19 Medisorb Technologies Internat LONG-TERM TREATMENT BY SLOW DELIVERY OF ANTISENSE-OLIGODEOXY-RIBONUCLEOTIDES FROM BIODEGRADABLE MICROPARTICLES
US5456986A (en) 1993-06-30 1995-10-10 Carnegie Mellon University Magnetic metal or metal carbide nanoparticles and a process for forming same
DE69429038T2 (de) 1993-07-28 2002-03-21 Pe Corporation (Ny), Norwalk Vorrichtung und Verfahren zur Nukleinsäurevervielfältigung
US5874239A (en) 1993-07-30 1999-02-23 Affymax Technologies N.V. Biotinylation of proteins
ATE256735T1 (de) 1993-08-30 2004-01-15 Promega Corp Zusammensetzungen und verfahren zur reinigung von nukleinsäuren
US5512131A (en) 1993-10-04 1996-04-30 President And Fellows Of Harvard College Formation of microstamped patterns on surfaces and derivative articles
US20030044777A1 (en) 1993-10-28 2003-03-06 Kenneth L. Beattie Flowthrough devices for multiple discrete binding reactions
US5605793A (en) 1994-02-17 1997-02-25 Affymax Technologies N.V. Methods for in vitro recombination
US5558071A (en) 1994-03-07 1996-09-24 Combustion Electromagnetics, Inc. Ignition system power converter and controller
US5648211A (en) 1994-04-18 1997-07-15 Becton, Dickinson And Company Strand displacement amplification using thermophilic enzymes
WO1995031726A1 (en) 1994-05-11 1995-11-23 Genera Technologies Limited Methods of capturing species from liquids and assay procedures
US5705628A (en) 1994-09-20 1998-01-06 Whitehead Institute For Biomedical Research DNA purification and isolation using magnetic particles
WO1998053300A2 (en) 1997-05-23 1998-11-26 Lynx Therapeutics, Inc. System and apparaus for sequential processing of analytes
US5846719A (en) 1994-10-13 1998-12-08 Lynx Therapeutics, Inc. Oligonucleotide tags for sorting and identification
US5585069A (en) 1994-11-10 1996-12-17 David Sarnoff Research Center, Inc. Partitioned microelectronic and fluidic device array for clinical diagnostics and chemical synthesis
EP0812434B1 (en) 1995-03-01 2013-09-18 President and Fellows of Harvard College Microcontact printing on surfaces and derivative articles
US5700642A (en) 1995-05-22 1997-12-23 Sri International Oligonucleotide sizing using immobilized cleavable primers
HUP9900910A2 (hu) 1995-06-07 1999-07-28 Lynx Therapeutics, Inc. Oligonukleotid jelzések osztályozáshoz és azonosításhoz
EP1967592B1 (en) 1995-06-07 2010-04-28 Solexa, Inc. Method of improving the efficiency of polynucleotide sequencing
US5856174A (en) 1995-06-29 1999-01-05 Affymetrix, Inc. Integrated nucleic acid diagnostic device
US6866760B2 (en) 1998-08-27 2005-03-15 E Ink Corporation Electrophoretic medium and process for the production thereof
US5872010A (en) 1995-07-21 1999-02-16 Northeastern University Microscale fluid handling system
US6057149A (en) 1995-09-15 2000-05-02 The University Of Michigan Microscale devices and reactions in microscale devices
US5851769A (en) 1995-09-27 1998-12-22 The Regents Of The University Of California Quantitative DNA fiber mapping
US5736330A (en) 1995-10-11 1998-04-07 Luminex Corporation Method and compositions for flow cytometric determination of DNA sequences
US5736332A (en) 1995-11-30 1998-04-07 Mandecki; Wlodek Method of determining the sequence of nucleic acids employing solid-phase particles carrying transponders
US6001571A (en) 1995-11-30 1999-12-14 Mandecki; Wlodek Multiplex assay for nucleic acids employing transponders
US6051377A (en) 1995-11-30 2000-04-18 Pharmaseq, Inc. Multiplex assay for nucleic acids employing transponders
US6355198B1 (en) 1996-03-15 2002-03-12 President And Fellows Of Harvard College Method of forming articles including waveguides via capillary micromolding and microtransfer molding
EP0832436A1 (en) 1996-04-15 1998-04-01 Dade Behring Inc. Apparatus and method for analysis
JP4468488B2 (ja) 1996-05-29 2010-05-26 コーネル・リサーチ・ファンデーション・インコーポレイテッド 組み合せたリガーゼ検出およびポリメラーゼ連鎖反応を用いる核酸配列相違の検出
US5846727A (en) 1996-06-06 1998-12-08 Board Of Supervisors Of Louisiana State University And Agricultural & Mechanical College Microsystem for rapid DNA sequencing
US6458335B1 (en) 1996-07-15 2002-10-01 Calcitech Ltd. Production of powders
US5965443A (en) 1996-09-09 1999-10-12 Wisconsin Alumni Research Foundation System for in vitro transposition
US6133436A (en) 1996-11-06 2000-10-17 Sequenom, Inc. Beads bound to a solid support and to nucleic acids
US5900481A (en) 1996-11-06 1999-05-04 Sequenom, Inc. Bead linkers for immobilizing nucleic acids to solid supports
AU746549B2 (en) 1996-11-20 2002-05-02 Becton Dickinson & Company Microfabricated isothermal nucleic acid amplification devices and methods
US5958703A (en) 1996-12-03 1999-09-28 Glaxo Group Limited Use of modified tethers in screening compound libraries
US20020172965A1 (en) 1996-12-13 2002-11-21 Arcaris, Inc. Methods for measuring relative amounts of nucleic acids in a complex mixture and retrieval of specific sequences therefrom
US20050042625A1 (en) 1997-01-15 2005-02-24 Xzillion Gmbh & Co. Mass label linked hybridisation probes
US6297006B1 (en) 1997-01-16 2001-10-02 Hyseq, Inc. Methods for sequencing repetitive sequences and for determining the order of sequence subfragments
US20020034737A1 (en) 1997-03-04 2002-03-21 Hyseq, Inc. Methods and compositions for detection or quantification of nucleic acid species
AU762888B2 (en) 1997-02-12 2003-07-10 Us Genomics Methods and products for analyzing polymers
US7622294B2 (en) 1997-03-14 2009-11-24 Trustees Of Tufts College Methods for detecting target analytes and enzymatic reactions
US6327410B1 (en) 1997-03-14 2001-12-04 The Trustees Of Tufts College Target analyte sensors utilizing Microspheres
US20030027203A1 (en) 1997-03-24 2003-02-06 Fields Robert E. Biomolecular processor
US6391622B1 (en) 1997-04-04 2002-05-21 Caliper Technologies Corp. Closed-loop biochemical analyzers
US6143496A (en) 1997-04-17 2000-11-07 Cytonix Corporation Method of sampling, amplifying and quantifying segment of nucleic acid, polymerase chain reaction assembly having nanoliter-sized sample chambers, and method of filling assembly
US6110678A (en) 1997-05-02 2000-08-29 Gen-Probe Incorporated Two-step hybridization and capture of a polynucleotide
DE69823347T2 (de) 1997-05-16 2005-05-12 Alberta Research Council, Edmonton Mikrofluidisches system und verfahren zu dessen betrieb
US6969488B2 (en) 1998-05-22 2005-11-29 Solexa, Inc. System and apparatus for sequential processing of analytes
US20040241759A1 (en) 1997-06-16 2004-12-02 Eileen Tozer High throughput screening of libraries
EP2258846A3 (en) 1997-07-07 2012-03-21 Medical Research Council A method for increasing the concentration of a nucleic acid molecule
GB9714716D0 (en) 1997-07-11 1997-09-17 Brax Genomics Ltd Characterising nucleic acids
FI103809B (fi) 1997-07-14 1999-09-30 Finnzymes Oy In vitro -menetelmä templaattien tuottamiseksi DNA-sekventointia varten
US6974669B2 (en) 2000-03-28 2005-12-13 Nanosphere, Inc. Bio-barcodes based on oligonucleotide-modified nanoparticles
US20050037397A1 (en) 2001-03-28 2005-02-17 Nanosphere, Inc. Bio-barcode based detection of target analytes
US6368871B1 (en) 1997-08-13 2002-04-09 Cepheid Non-planar microstructures for manipulation of fluid samples
AU8908198A (en) 1997-08-15 1999-03-08 Hyseq, Inc. Methods and compositions for detection or quantification of nucleic acid species
WO1999014368A2 (en) 1997-09-15 1999-03-25 Whitehead Institute For Biomedical Research Methods and apparatus for processing a sample of biomolecular analyte using a microfabricated device
US20020092767A1 (en) 1997-09-19 2002-07-18 Aclara Biosciences, Inc. Multiple array microfluidic device units
US7214298B2 (en) 1997-09-23 2007-05-08 California Institute Of Technology Microfabricated cell sorter
US6103537A (en) 1997-10-02 2000-08-15 Aclara Biosciences, Inc. Capillary assays involving separation of free and bound species
US5842787A (en) 1997-10-09 1998-12-01 Caliper Technologies Corporation Microfluidic systems incorporating varied channel dimensions
US6511803B1 (en) 1997-10-10 2003-01-28 President And Fellows Of Harvard College Replica amplification of nucleic acid arrays
CA2305449A1 (en) 1997-10-10 1999-04-22 President & Fellows Of Harvard College Replica amplification of nucleic acid arrays
US6485944B1 (en) 1997-10-10 2002-11-26 President And Fellows Of Harvard College Replica amplification of nucleic acid arrays
KR20010031140A (ko) 1997-10-14 2001-04-16 루미넥스 코포레이션 정밀 형광염료 입자 및 그의 제조방법 그리고 그의 사용
CA2306791C (en) 1997-10-24 2007-05-15 Northeastern University A multichannel microscale system for high throughput preparative separation with comprehensive collection and analysis
US6913935B1 (en) 1997-12-04 2005-07-05 Amersham Biosciences Uk Limited Multiple assay method
AU1726199A (en) 1997-12-31 1999-07-19 Chiron Corporation Metastatic cancer regulated gene
AU765378B2 (en) 1998-02-19 2003-09-18 President And Fellows Of Harvard College Monovalent, multivalent, and multimeric MHC binding domain fusion proteins and conjugates, and uses therefor
WO1999050402A1 (en) 1998-03-27 1999-10-07 President And Fellows Of Harvard College Systematic identification of essential genes by in vitro transposon mutagenesis
US6022716A (en) 1998-04-10 2000-02-08 Genset Sa High throughput DNA sequencing vector
AU3555599A (en) 1998-04-13 1999-11-01 Luminex Corporation Liquid labeling with fluorescent microparticles
US6048736A (en) 1998-04-29 2000-04-11 Kosak; Kenneth M. Cyclodextrin polymers for carrying and releasing drugs
US6780591B2 (en) 1998-05-01 2004-08-24 Arizona Board Of Regents Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
US5997636A (en) 1998-05-01 1999-12-07 Instrumentation Technology Associates, Inc. Method and apparatus for growing crystals
US6123798A (en) 1998-05-06 2000-09-26 Caliper Technologies Corp. Methods of fabricating polymeric structures incorporating microscale fluidic elements
WO1999058664A1 (en) 1998-05-14 1999-11-18 Whitehead Institute For Biomedical Research Solid phase technique for selectively isolating nucleic acids
US6306590B1 (en) 1998-06-08 2001-10-23 Caliper Technologies Corp. Microfluidic matrix localization apparatus and methods
CA2335951C (en) 1998-06-24 2013-07-30 Mark S. Chee Decoding of array sensors with microspheres
WO2000008212A1 (en) 1998-08-07 2000-02-17 Cellay, Llc Gel microdrops in genetic analysis
US6159736A (en) 1998-09-23 2000-12-12 Wisconsin Alumni Research Foundation Method for making insertional mutations using a Tn5 synaptic complex
AR021833A1 (es) 1998-09-30 2002-08-07 Applied Research Systems Metodos de amplificacion y secuenciacion de acido nucleico
WO2000022436A1 (en) 1998-10-13 2000-04-20 Biomicro Systems, Inc. Fluid circuit components based upon passive fluid dynamics
US6489096B1 (en) 1998-10-15 2002-12-03 Princeton University Quantitative analysis of hybridization patterns and intensities in oligonucleotide arrays
SE9803614L (sv) 1998-10-19 2000-04-20 Muhammed Mamoun Förfarande och anordning för framställning av nanopartiklar
WO2000026412A1 (en) 1998-11-02 2000-05-11 Kenneth Loren Beattie Nucleic acid analysis using sequence-targeted tandem hybridization
US6569631B1 (en) 1998-11-12 2003-05-27 3-Dimensional Pharmaceuticals, Inc. Microplate thermal shift assay for ligand development using 5-(4″dimethylaminophenyl)-2-(4′-phenyl)oxazole derivative fluorescent dyes
US5942609A (en) 1998-11-12 1999-08-24 The Porkin-Elmer Corporation Ligation assembly and detection of polynucleotides on solid-support
WO2000034527A2 (en) 1998-12-11 2000-06-15 The Regents Of The University Of California Targeted molecular bar codes
NO986133D0 (no) 1998-12-23 1998-12-23 Preben Lexow FremgangsmÕte for DNA-sekvensering
GB9900298D0 (en) 1999-01-07 1999-02-24 Medical Res Council Optical sorting method
US6416642B1 (en) 1999-01-21 2002-07-09 Caliper Technologies Corp. Method and apparatus for continuous liquid flow in microscale channels using pressure injection, wicking, and electrokinetic injection
US6635419B1 (en) 1999-02-16 2003-10-21 Applera Corporation Polynucleotide sequencing method
US20030027214A1 (en) 1999-02-17 2003-02-06 Kamb Carl Alexander Methods for substrate-ligand interaction screening
CA2362263A1 (en) * 1999-02-19 2000-08-24 Bioserv Ag Biodegradable composite material for the production of microcapsules
EP1163052B1 (en) 1999-02-23 2010-06-02 Caliper Life Sciences, Inc. Manipulation of microparticles in microfluidic systems
US7615373B2 (en) 1999-02-25 2009-11-10 Virginia Commonwealth University Intellectual Property Foundation Electroprocessed collagen and tissue engineering
US6171850B1 (en) 1999-03-08 2001-01-09 Caliper Technologies Corp. Integrated devices and systems for performing temperature controlled reactions and analyses
US6303343B1 (en) 1999-04-06 2001-10-16 Caliper Technologies Corp. Inefficient fast PCR
US6908737B2 (en) 1999-04-15 2005-06-21 Vitra Bioscience, Inc. Systems and methods of conducting multiplexed experiments
US20060275782A1 (en) 1999-04-20 2006-12-07 Illumina, Inc. Detection of nucleic acid reactions on bead arrays
ATE553219T1 (de) 1999-04-20 2012-04-15 Illumina Inc Erkennung von nukleinsäurereaktionen auf bead- arrays
JP2002542781A (ja) 1999-04-28 2002-12-17 ザ、ボード、オブ、トラスティーズ、オブ、ザ、リーランド、スタンフォード、ジュニア、ユニバーシティ Ｐ因子由来ベクター及びその使用法
US6399952B1 (en) 1999-05-12 2002-06-04 Aclara Biosciences, Inc. Multiplexed fluorescent detection in microfluidic devices
WO2000070095A2 (en) 1999-05-17 2000-11-23 Dade Behring Inc. Homogeneous isothermal amplification and detection of nucleic acids using a template switch oligonucleotide
US20020051971A1 (en) 1999-05-21 2002-05-02 John R. Stuelpnagel Use of microfluidic systems in the detection of target analytes using microsphere arrays
US6846622B1 (en) 1999-05-26 2005-01-25 Oregon Health & Science University Tagged epitope protein transposable element
US20030124509A1 (en) 1999-06-03 2003-07-03 Kenis Paul J.A. Laminar flow patterning and articles made thereby
US7098032B2 (en) 2001-01-02 2006-08-29 Mirus Bio Corporation Compositions and methods for drug delivery using pH sensitive molecules
US6372813B1 (en) 1999-06-25 2002-04-16 Motorola Methods and compositions for attachment of biomolecules to solid supports, hydrogels, and hydrogel arrays
AU6068300A (en) 1999-07-06 2001-01-22 Caliper Technologies Corporation Microfluidic systems and methods for determining modulator kinetics
US6977145B2 (en) 1999-07-28 2005-12-20 Serono Genetics Institute S.A. Method for carrying out a biochemical protocol in continuous flow in a microreactor
US6524456B1 (en) 1999-08-12 2003-02-25 Ut-Battelle, Llc Microfluidic devices for the controlled manipulation of small volumes
EP1210358A4 (en) 1999-08-13 2005-01-05 Univ Brandeis DETECTION OF NUCLEIC ACIDS
EP1248853A2 (en) 1999-08-20 2002-10-16 Luminex Corporation Liquid array technology
JP2003508763A (ja) 1999-08-27 2003-03-04 マトリックス テクノロジーズ コーポレイション 固体支持体上にリガンドを固定化する方法及び装置並びにその使用方法
US6982146B1 (en) 1999-08-30 2006-01-03 The United States Of America As Represented By The Department Of Health And Human Services High speed parallel molecular nucleic acid sequencing
IL149001A0 (en) 1999-10-13 2002-11-10 Signature Bioscience Inc System and method for detecting and identifying molecular events in test sample
US6958225B2 (en) 1999-10-27 2005-10-25 Affymetrix, Inc. Complexity management of genomic DNA
AU1100201A (en) 1999-10-28 2001-05-08 Board Of Trustees Of The Leland Stanford Junior University Methods of in vivo gene transfer using a sleeping beauty transposon system
HK1052203A1 (zh) 1999-11-08 2003-09-05 荣研化学株式会社 检测突变和/或多态性的方法
US6432290B1 (en) 1999-11-26 2002-08-13 The Governors Of The University Of Alberta Apparatus and method for trapping bead based reagents within microfluidic analysis systems
US20010051348A1 (en) 2000-01-28 2001-12-13 Lee Chee Wee Novel ligands and methods for preparing same
WO2001063241A2 (en) 2000-02-23 2001-08-30 Zyomyx, Inc. Microfluidic devices and methods
MXPA02008290A (es) 2000-02-23 2004-09-10 Caliper Technologies Inc Sistema de control de presion de multi-reserva-.
IL134830A0 (en) 2000-03-01 2001-05-20 Chay 13 Medical Res Group N V Peptides and immunostimulatory and anti-bacterial pharmaceutical compositions containing them
CN1487836A (zh) 2000-03-14 2004-04-07 埃米林药品公司 胰高糖素样肽1(7-36)对胃窦幽门十二指肠能动性的影响
US6376191B1 (en) * 2000-03-22 2002-04-23 Mergen, Ltd. Microarray-based analysis of polynucleotide sequence variations
US6409832B2 (en) 2000-03-31 2002-06-25 Micronics, Inc. Protein crystallization in microfluidic structures
US20020001856A1 (en) 2000-04-06 2002-01-03 Chow Andrea W. Methods and devices for achieving long incubation times in high-throughput systems
EP1313879A2 (en) 2000-04-10 2003-05-28 Matthew Ashby Methods for the survey and genetic analysis of populations
US6481453B1 (en) 2000-04-14 2002-11-19 Nanostream, Inc. Microfluidic branch metering systems and methods
US6800298B1 (en) 2000-05-11 2004-10-05 Clemson University Biological lubricant composition and method of applying lubricant composition
US20060008799A1 (en) 2000-05-22 2006-01-12 Hong Cai Rapid haplotyping by single molecule detection
WO2001089675A2 (en) 2000-05-24 2001-11-29 Micronics, Inc. Jet vortex mixer
US6645432B1 (en) 2000-05-25 2003-11-11 President & Fellows Of Harvard College Microfluidic systems including three-dimensionally arrayed channel networks
US20060263888A1 (en) 2000-06-02 2006-11-23 Honeywell International Inc. Differential white blood count on a disposable card
US6632606B1 (en) 2000-06-12 2003-10-14 Aclara Biosciences, Inc. Methods for single nucleotide polymorphism detection
AU7299301A (en) 2000-06-21 2002-01-02 Bioarray Solutions Ltd Multianalyte molecular analysis using application-specific random particle arrays
AU2001281076A1 (en) 2000-08-07 2002-02-18 Nanostream, Inc. Fluidic mixer in microfluidic system
US6610499B1 (en) 2000-08-31 2003-08-26 The Regents Of The University Of California Capillary array and related methods
US6773566B2 (en) 2000-08-31 2004-08-10 Nanolytics, Inc. Electrostatic actuators for microfluidics and methods for using same
ATE448875T1 (de) 2000-09-14 2009-12-15 Caliper Life Sciences Inc Mikrofluidische vorrichtungen und methoden um temperatur-vermittelte reaktionen durchzuführen
EP2299256A3 (en) 2000-09-15 2012-10-10 California Institute Of Technology Microfabricated crossflow devices and methods
US7258774B2 (en) 2000-10-03 2007-08-21 California Institute Of Technology Microfluidic devices and methods of use
WO2002031203A2 (en) 2000-10-10 2002-04-18 Diversa Corporation High throughput or capillary-based screening for a bioactivity or biomolecule
US7045283B2 (en) 2000-10-18 2006-05-16 The Regents Of The University Of California Methods of high-throughput screening for internalizing antibodies
JP2002155305A (ja) 2000-11-14 2002-05-31 Akira Kawasaki 単分散粒子の製造装置及び単分散粒子の製造方法及びその製造方法で製造された単分散粒子
US6492154B2 (en) 2001-01-31 2002-12-10 Applera Corporation Isolated human kinase proteins, nucleic acid molecules encoding human kinase proteins, and uses thereof
CA2332186A1 (en) 2001-02-08 2002-08-08 Her Majesty In Right Of Canada As Represented By The Minister Of Agricul Ture And Agri-Food Canada Replicative in vivo gene targeting
US7670559B2 (en) 2001-02-15 2010-03-02 Caliper Life Sciences, Inc. Microfluidic systems with enhanced detection systems
DE60207831D1 (de) * 2001-02-22 2006-01-12 Anika Therapeutics Inc Thiol-modifizierte hyaluronan-derivate
EP1362634B1 (en) 2001-02-23 2006-05-31 Japan Science and Technology Agency Process for producing emulsion and apparatus therefor
US7211654B2 (en) 2001-03-14 2007-05-01 Regents Of The University Of Michigan Linkers and co-coupling agents for optimization of oligonucleotide synthesis and purification on solid supports
US20150329617A1 (en) 2001-03-14 2015-11-19 Dynal Biotech Asa Novel MHC molecule constructs, and methods of employing these constructs for diagnosis and therapy, and uses of MHC molecules
US20060040286A1 (en) 2001-03-28 2006-02-23 Nanosphere, Inc. Bio-barcode based detection of target analytes
EP1377821A2 (en) 2001-04-03 2004-01-07 Micronics, Inc. Pneumatic valve interface for use in microfluidic structures
US7138267B1 (en) 2001-04-04 2006-11-21 Epicentre Technologies Corporation Methods and compositions for amplifying DNA clone copy number
US20030027221A1 (en) * 2001-04-06 2003-02-06 Scott Melissa E. High-throughput screening assays by encapsulation
US7572642B2 (en) 2001-04-18 2009-08-11 Ambrigen, Llc Assay based on particles, which specifically bind with targets in spatially distributed characteristic patterns
US6806058B2 (en) 2001-05-26 2004-10-19 One Cell Systems, Inc. Secretions of proteins by encapsulated cells
US6880576B2 (en) 2001-06-07 2005-04-19 Nanostream, Inc. Microfluidic devices for methods development
US6524259B2 (en) * 2001-06-08 2003-02-25 Cervilenz, Inc. Devices and methods for cervix measurement
US7179423B2 (en) 2001-06-20 2007-02-20 Cytonome, Inc. Microfluidic system including a virtual wall fluid interface port for interfacing fluids with the microfluidic system
US7262063B2 (en) 2001-06-21 2007-08-28 Bio Array Solutions, Ltd. Directed assembly of functional heterostructures
US6613523B2 (en) 2001-06-29 2003-09-02 Agilent Technologies, Inc. Method of DNA sequencing using cleavable tags
US7682353B2 (en) 2001-06-29 2010-03-23 Coloplast A/S Catheter device
US7077152B2 (en) 2001-07-07 2006-07-18 Nanostream, Inc. Microfluidic metering systems and methods
IL159957A0 (en) 2001-07-20 2004-06-20 California Inst Of Techn Protein and nucleic expression systems
US6767731B2 (en) 2001-08-27 2004-07-27 Intel Corporation Electron induced fluorescent method for nucleic acid sequencing
AU2002360272A1 (en) 2001-10-10 2003-04-22 Superarray Bioscience Corporation Detecting targets by unique identifier nucleotide tags
US7297485B2 (en) 2001-10-15 2007-11-20 Qiagen Gmbh Method for nucleic acid amplification that results in low amplification bias
US6783647B2 (en) 2001-10-19 2004-08-31 Ut-Battelle, Llc Microfluidic systems and methods of transport and lysis of cells and analysis of cell lysate
AU2002360282A1 (en) 2001-10-19 2003-06-10 West Virginia University Research Corporation Microflludic system for proteome analysis
US20030149307A1 (en) 2001-10-24 2003-08-07 Baxter International Inc. Process for the preparation of polyethylene glycol bis amine
JP2005514224A (ja) 2001-10-26 2005-05-19 アクララ バイオサイエンシーズ， インコーポレイテッド ミクロ流体基材の射出成形ミクロ複製のためのシステムおよび方法
CA2466164A1 (en) 2001-10-30 2003-05-08 Nanomics Biosystems Pty, Ltd. Device and methods for directed synthesis of chemical libraries
US7262056B2 (en) 2001-11-08 2007-08-28 Mirus Bio Corporation Enhancing intermolecular integration of nucleic acids using integrator complexes
GB0127564D0 (en) 2001-11-16 2002-01-09 Medical Res Council Emulsion compositions
AU2002351193A1 (en) 2001-11-28 2003-06-10 Mj Bioworks Incorporated Parallel polymorphism scoring by amplification and error correction
US7335153B2 (en) 2001-12-28 2008-02-26 Bio Array Solutions Ltd. Arrays of microparticles and methods of preparation thereof
WO2003057010A2 (en) 2002-01-04 2003-07-17 Board Of Regents, The University Of Texas System Droplet-based microfluidic oligonucleotide synthesis engine
CA2473376A1 (en) 2002-01-16 2003-07-31 Dynal Biotech Asa Method for isolating nucleic acids and protein from a single sample
KR100459870B1 (ko) 2002-02-22 2004-12-04 한국과학기술원 트랜스포존과 Cre/loxP 부위 특이적 재조합 방법을 이용하는 염색체의 특정부위가 제거된 미생물 변이주 제조방법
EP1488006B1 (en) 2002-03-20 2008-05-28 InnovativeBio.Biz Microcapsules with controlable permeability encapsulating a nucleic acid amplification reaction mixture and their use as reaction compartments for parallels reactions
EP2278338B1 (en) 2002-05-09 2020-08-26 The University of Chicago Device and method for pressure-driven plug transport and reaction
US7901939B2 (en) 2002-05-09 2011-03-08 University Of Chicago Method for performing crystallization and reactions in pressure-driven fluid plugs
US7527966B2 (en) 2002-06-26 2009-05-05 Transgenrx, Inc. Gene regulation in transgenic animals using a transposon-based vector
JP2006507921A (ja) 2002-06-28 2006-03-09 プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ 流体分散のための方法および装置
IL151660A0 (en) 2002-09-09 2003-04-10 Univ Ben Gurion Method for isolating and culturing unculturable microorganisms
EP1546376B9 (en) 2002-09-30 2009-08-12 F.Hoffmann-La Roche Ag Oligonucleotides for genotyping thymidylate synthase gene
US20040081962A1 (en) 2002-10-23 2004-04-29 Caifu Chen Methods for synthesizing complementary DNA
US6979713B2 (en) 2002-11-25 2005-12-27 3M Innovative Properties Company Curable compositions and abrasive articles therefrom
US20050266582A1 (en) 2002-12-16 2005-12-01 Modlin Douglas N Microfluidic system with integrated permeable membrane
US20040248299A1 (en) 2002-12-27 2004-12-09 Sumedha Jayasena RNA interference
WO2004065617A2 (en) 2003-01-17 2004-08-05 The Trustees Of Boston University Haplotype analysis
AU2004209416B2 (en) 2003-01-29 2007-08-02 454 Life Sciences Corporation Double ended sequencing
WO2004070042A1 (en) 2003-02-10 2004-08-19 Max-Delbrück-Centrum Für Molekulare Medizin (Mdc) Transposon-based targeting system
US7041481B2 (en) 2003-03-14 2006-05-09 The Regents Of The University Of California Chemical amplification based on fluid partitioning
US7316903B2 (en) 2003-03-28 2008-01-08 United States Of America As Represented By The Department Of Health And Human Services Detection of nucleic acid sequence variations using phase Mu transposase
US20060078893A1 (en) 2004-10-12 2006-04-13 Medical Research Council Compartmentalised combinatorial chemistry by microfluidic control
GB0307428D0 (en) 2003-03-31 2003-05-07 Medical Res Council Compartmentalised combinatorial chemistry
GB0307403D0 (en) 2003-03-31 2003-05-07 Medical Res Council Selection by compartmentalised screening
RU2005130646A (ru) 2003-04-04 2006-01-27 Пфайзер Продактс Инк. (Us) Микрофлюидизированные эмульсии масло в воде и композиции вакцины
US20100035254A1 (en) 2003-04-08 2010-02-11 Pacific Biosciences Of California, Inc. Composition and method for nucleic acid sequencing
EP2127736A1 (en) 2003-04-10 2009-12-02 The President and Fellows of Harvard College Formation and control of fluidic species
EP1629286A1 (en) 2003-05-16 2006-03-01 Global Technologies (NZ) Ltd. Method and apparatus for mixing sample and reagent in a suspension fluid
WO2004103565A2 (de) 2003-05-19 2004-12-02 Hans-Knöll-Institut für Naturstoff-Forschung e.V. Vorrichtung und verfahren zur strukturierung von flüssigkeiten und zum zudosieren von reaktionsflüssigkeiten zu in separationsmedium eingebetteten flüssigkeitskompartimenten
WO2004105734A1 (en) 2003-05-28 2004-12-09 Valorisation Recherche, Societe En Commandite Method of preparing microcapsules
GB0313170D0 (en) 2003-06-09 2003-07-16 Qinetiq Ltd Method and apparatus for spore disruption and/or detection
WO2004113877A1 (en) 2003-06-13 2004-12-29 The General Hospital Corporation Microfluidic systems for size based removal of red blood cells and platelets from blood
CN101001960A (zh) * 2003-06-27 2007-07-18 西北大学 基于生物条形码检测靶分析物
GB2403475B (en) 2003-07-01 2008-02-06 Oxitec Ltd Stable integrands
GB0315438D0 (en) 2003-07-02 2003-08-06 Univ Manchester Analysis of mixed cell populations
JP5183063B2 (ja) 2003-07-05 2013-04-17 ザ ジョンズ ホプキンス ユニバーシティ 遺伝的変異の検出および列挙のための方法ならびに組成物
BRPI0414004A (pt) 2003-08-27 2006-10-24 Harvard College controle eletrÈnico de espécies fluìdicas
EP1660153A2 (en) 2003-09-04 2006-05-31 The United States of America as represented by the Department of Veterans Affairs Hydrogel nanocompsites for ophthalmic applications
EP1663497B2 (en) 2003-09-05 2020-03-25 Stokes Bio Limited A microfluidic analysis system
US7354706B2 (en) 2003-09-09 2008-04-08 The Regents Of The University Of Colorado, A Body Corporate Use of photopolymerization for amplification and detection of a molecular recognition event
JP4353945B2 (ja) 2003-09-22 2009-10-28 独立行政法人理化学研究所 効率的なｄｎａ逆位反復構造の調製方法
WO2005069001A1 (ja) 2003-09-25 2005-07-28 Toyama Prefecture マイクロウェルアレイチップ及びその製造方法
EP1694869A2 (en) 2003-11-10 2006-08-30 Investigen, Inc. Methods of preparing nucleic acid for detection
EP1691792A4 (en) 2003-11-24 2008-05-28 Yeda Res & Dev COMPOSITIONS AND PROCESSES FOR SORTING IN VITRO OF MOLECULE AND CELL LIBRARIES
US20050136417A1 (en) 2003-12-19 2005-06-23 Affymetrix, Inc. Amplification of nucleic acids
US8071364B2 (en) 2003-12-24 2011-12-06 Transgenrx, Inc. Gene therapy using transposon-based vectors
WO2005073410A2 (en) 2004-01-28 2005-08-11 454 Corporation Nucleic acid amplification with continuous flow emulsion
US20050181379A1 (en) 2004-02-18 2005-08-18 Intel Corporation Method and device for isolating and positioning single nucleic acid molecules
US20100216153A1 (en) 2004-02-27 2010-08-26 Helicos Biosciences Corporation Methods for detecting fetal nucleic acids and diagnosing fetal abnormalities
CA2557841A1 (en) 2004-02-27 2005-09-09 President And Fellows Of Harvard College Polony fluorescent in situ sequencing beads
KR100552706B1 (ko) 2004-03-12 2006-02-20 삼성전자주식회사 핵산 증폭 방법 및 장치
WO2005089921A1 (ja) 2004-03-23 2005-09-29 Japan Science And Technology Agency 微小液滴の生成方法及び装置
US20050221339A1 (en) 2004-03-31 2005-10-06 Medical Research Council Harvard University Compartmentalised screening by microfluidic control
US20060020371A1 (en) 2004-04-13 2006-01-26 President And Fellows Of Harvard College Methods and apparatus for manipulation and/or detection of biological samples and other objects
US20050250147A1 (en) 2004-05-10 2005-11-10 Macevicz Stephen C Digital profiling of polynucleotide populations
WO2006047183A2 (en) 2004-10-21 2006-05-04 New England Biolabs, Inc. Recombinant dna nicking endonuclease and uses thereof
US7799553B2 (en) 2004-06-01 2010-09-21 The Regents Of The University Of California Microfabricated integrated DNA analysis system
US7700281B2 (en) 2004-06-30 2010-04-20 Usb Corporation Hot start nucleic acid amplification
US7968085B2 (en) 2004-07-05 2011-06-28 Ascendis Pharma A/S Hydrogel formulations
CN1648671B (zh) 2005-02-06 2012-09-26 成都夸常医学工业有限公司 多反应器分析芯片检测方法和分析芯片及检测装置
JP4189760B2 (ja) * 2004-07-28 2008-12-03 アイシン精機株式会社 リクライニング装置
US7608434B2 (en) 2004-08-04 2009-10-27 Wisconsin Alumni Research Foundation Mutated Tn5 transposase proteins and the use thereof
US7892731B2 (en) 2004-10-01 2011-02-22 Radix Biosolutions, Ltd. System and method for inhibiting the decryption of a nucleic acid probe sequence used for the detection of a specific nucleic acid
US7968287B2 (en) 2004-10-08 2011-06-28 Medical Research Council Harvard University In vitro evolution in microfluidic systems
US9492400B2 (en) 2004-11-04 2016-11-15 Massachusetts Institute Of Technology Coated controlled release polymer particles as efficient oral delivery vehicles for biopharmaceuticals
US20080004436A1 (en) 2004-11-15 2008-01-03 Yeda Research And Development Co. Ltd. At The Weizmann Institute Of Science Directed Evolution and Selection Using in Vitro Compartmentalization
US7329493B2 (en) 2004-12-22 2008-02-12 Asiagen Corporation One-tube nested PCR for detecting Mycobacterium tuberculosis
US8883487B2 (en) 2004-12-23 2014-11-11 Abbott Point Of Care Inc. Molecular diagnostics system and methods
US20080213593A1 (en) 2005-01-21 2008-09-04 President And Fellows Of Harvard College Systems And Methods For Forming Fluidic Droplets Encapsulated In Particles Such As Colloidal Particles
EP1841879A4 (en) 2005-01-25 2009-05-27 Population Genetics Technologi ISOTHERMAL DNA AMPLIFICATION
US7407757B2 (en) 2005-02-10 2008-08-05 Population Genetics Technologies Genetic analysis by sequence-specific sorting
US7393665B2 (en) 2005-02-10 2008-07-01 Population Genetics Technologies Ltd Methods and compositions for tagging and identifying polynucleotides
AU2006214085A1 (en) 2005-02-18 2006-08-24 Canon U.S. Life Sciences, Inc. Devices and methods for identifying genomic DNA of organisms
EP1867702B1 (en) 2005-02-21 2011-09-28 Kagoshima University Method for purifying biodiesel fuel
US20070054119A1 (en) 2005-03-04 2007-03-08 Piotr Garstecki Systems and methods of forming particles
US9040237B2 (en) 2005-03-04 2015-05-26 Intel Corporation Sensor arrays and nucleic acid sequencing applications
CA2599683A1 (en) 2005-03-04 2006-09-14 President And Fellows Of Harvard College Method and apparatus for forming multiple emulsions
JP2006289250A (ja) 2005-04-08 2006-10-26 Kao Corp マイクロミキサー及びそれを用いた流体混合方法
US8407013B2 (en) 2005-06-07 2013-03-26 Peter K. Rogan AB initio generation of single copy genomic probes
EP2620510B2 (en) 2005-06-15 2020-02-19 Complete Genomics Inc. Single molecule arrays for genetic and chemical analysis
JP2006349060A (ja) 2005-06-16 2006-12-28 Ntn Corp ボールねじ
US8828209B2 (en) 2005-06-22 2014-09-09 The Research Foundation For The State University Of New York Massively parallel 2-dimensional capillary electrophoresis
WO2007002567A2 (en) 2005-06-23 2007-01-04 Nanosphere, Inc. Selective isolation and concentration of nucleic acids from complex samples
EP1921140B1 (en) 2005-07-05 2011-12-14 Juridical Foundation The Chemo-Sero-Therapeutic Research Institute Mutant transposon vector and use thereof
JP5051490B2 (ja) * 2005-07-08 2012-10-17 独立行政法人産業技術総合研究所 マクロ生体材料を内包する無機マイクロカプセルおよびその製造方法
US20070020640A1 (en) 2005-07-21 2007-01-25 Mccloskey Megan L Molecular encoding of nucleic acid templates for PCR and other forms of sequence analysis
FR2888912B1 (fr) 2005-07-25 2007-08-24 Commissariat Energie Atomique Procede de commande d'une communication entre deux zones par electromouillage, dispositif comportant des zones isolables les unes des autres et procede de realisation d'un tel dispositif
WO2007018601A1 (en) 2005-08-02 2007-02-15 Rubicon Genomics, Inc. Compositions and methods for processing and amplification of dna, including using multiple enzymes in a single reaction
DE102005037401B4 (de) 2005-08-08 2007-09-27 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Bildung einer Emulsion in einem fluidischen Mikrosystem
WO2007024840A2 (en) 2005-08-22 2007-03-01 Critical Therapeutics, Inc. Method of quantitating nucleic acids by flow cytometry microparticle-based array
US7556776B2 (en) 2005-09-08 2009-07-07 President And Fellows Of Harvard College Microfluidic manipulation of fluids and reactions
JP2007074967A (ja) 2005-09-13 2007-03-29 Canon Inc 識別子プローブ及びそれを用いた核酸増幅方法
JP2009513948A (ja) * 2005-09-16 2009-04-02 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 検体検出のための比色バイオバーコード増幅アッセイ
US7960104B2 (en) 2005-10-07 2011-06-14 Callida Genomics, Inc. Self-assembled single molecule arrays and uses thereof
WO2007133831A2 (en) 2006-02-24 2007-11-22 Callida Genomics, Inc. High throughput genome sequencing on dna arrays
US20070111241A1 (en) 2005-10-14 2007-05-17 Nezih Cereb System and method for accessing, tracking, and editing sequence analysis and software to accomplish the same
US7709544B2 (en) 2005-10-25 2010-05-04 Massachusetts Institute Of Technology Microstructure synthesis by flow lithography and polymerization
EP1954838B1 (en) 2005-11-14 2014-02-26 Life Technologies Corporation Coded molecules for detecting target analytes
US20070134277A1 (en) 2005-12-09 2007-06-14 Children's Medical Center Corporation Pharmaceutical formulation for sulfur-containing drugs in liquid dosage forms
US20070141584A1 (en) 2005-12-20 2007-06-21 Roberts Douglas N Methods for assessment of native chromatin on microarrays
US7932037B2 (en) 2007-12-05 2011-04-26 Perkinelmer Health Sciences, Inc. DNA assays using amplicon probes on encoded particles
WO2007081385A2 (en) 2006-01-11 2007-07-19 Raindance Technologies, Inc. Microfluidic devices and methods of use in the formation and control of nanoreactors
WO2007087310A2 (en) 2006-01-23 2007-08-02 Population Genetics Technologies Ltd. Nucleic acid analysis using sequence tokens
WO2007087312A2 (en) 2006-01-23 2007-08-02 Population Genetics Technologies Ltd. Molecular counting
CA2640024A1 (en) 2006-01-27 2007-08-09 President And Fellows Of Harvard College Fluidic droplet coalescence
ES2429408T5 (es) 2006-02-02 2020-01-16 Univ Leland Stanford Junior Examen genético fetal no invasivo mediante análisis digital
WO2007092538A2 (en) 2006-02-07 2007-08-16 President And Fellows Of Harvard College Methods for making nucleotide probes for sequencing and synthesis
GB0603251D0 (en) 2006-02-17 2006-03-29 Isis Innovation DNA conformation
SG10201405158QA (en) 2006-02-24 2014-10-30 Callida Genomics Inc High throughput genome sequencing on dna arrays
WO2007111937A1 (en) 2006-03-23 2007-10-04 Applera Corporation Directed enrichment of genomic dna for high-throughput sequencing
JP4921829B2 (ja) 2006-03-30 2012-04-25 株式会社東芝 微粒子の製造装置、乳化剤保持部、微粒子の製造方法および分子膜の製造方法
WO2007114794A1 (en) 2006-03-31 2007-10-11 Nam Trung Nguyen Active control for droplet-based microfluidics
US20090062129A1 (en) 2006-04-19 2009-03-05 Agencourt Personal Genomics, Inc. Reagents, methods, and libraries for gel-free bead-based sequencing
US7811603B2 (en) 2006-05-09 2010-10-12 The Regents Of The University Of California Microfluidic device for forming monodisperse lipoplexes
EP2047910B1 (en) 2006-05-11 2012-01-11 Raindance Technologies, Inc. Microfluidic device and method
ES2620398T3 (es) 2006-05-22 2017-06-28 Nanostring Technologies, Inc. Sistemas y métodos para analizar nanoindicadores
RU2321638C2 (ru) 2006-05-23 2008-04-10 Закрытое акционерное общество "Молекулярно-медицинские технологии" Способ изготовления многофункционального мультичипа, мультичип для последовательного или параллельного скрининга биополимеров, способ анализа биополимеров и набор для осуществления способа
CA2653321A1 (en) 2006-05-26 2007-12-06 Althea Technologies, Inc. Biochemical analysis of partitioned cells
FR2901717A1 (fr) 2006-05-30 2007-12-07 Centre Nat Rech Scient Procede de traitement de gouttes dans un circuit microfluidique.
AU2007260676A1 (en) 2006-06-14 2007-12-21 Artemis Health, Inc. Rare cell analysis using sample splitting and DNA tags
WO2007149432A2 (en) 2006-06-19 2007-12-27 The Johns Hopkins University Single-molecule pcr on microparticles in water-in-oil emulsions
US20080076909A1 (en) 2006-06-30 2008-03-27 Applera Corporation Emulsion pcr and amplicon capture
EP1878501A1 (en) 2006-07-14 2008-01-16 Roche Diagnostics GmbH Instrument for heating and cooling
US8394590B2 (en) 2006-08-02 2013-03-12 California Institute Of Technology Capture agents and related methods and systems for detecting and/or sorting targets
US9012390B2 (en) 2006-08-07 2015-04-21 Raindance Technologies, Inc. Fluorocarbon emulsion stabilizing surfactants
JP5553602B2 (ja) 2006-09-06 2014-07-16 キヤノン ユー．エス． ライフ サイエンシズ， インコーポレイテッド マイクロ流体アッセイを実施するためのチップ及びカートリッジ設計構成
EP2066699B1 (en) 2006-09-25 2020-02-12 Archer-Daniels-Midland Company Superabsorbent surface-treated carboxyalkylated polysaccharides and process for producing same
US7935518B2 (en) 2006-09-27 2011-05-03 Alessandra Luchini Smart hydrogel particles for biomarker harvesting
US20080166720A1 (en) 2006-10-06 2008-07-10 The Regents Of The University Of California Method and apparatus for rapid nucleic acid analysis
US8841116B2 (en) 2006-10-25 2014-09-23 The Regents Of The University Of California Inline-injection microdevice and microfabricated integrated DNA analysis system using same
US7910354B2 (en) 2006-10-27 2011-03-22 Complete Genomics, Inc. Efficient arrays of amplified polynucleotides
US8709787B2 (en) 2006-11-14 2014-04-29 Handylab, Inc. Microfluidic cartridge and method of using same
CA2669728C (en) 2006-11-15 2017-04-11 Biospherex Llc Multitag sequencing and ecogenomics analysis
KR20090106484A (ko) 2006-11-20 2009-10-09 나티비스, 인코포레이티드 저주파 신호로 시험관내 또는 포유류 시스템을 변환시키기 위한 장치 및 방법
US20080242560A1 (en) * 2006-11-21 2008-10-02 Gunderson Kevin L Methods for generating amplified nucleic acid arrays
US8598328B2 (en) 2006-12-13 2013-12-03 National University Corporation Nagoya University Tol1 factor transposase and DNA introduction system using the same
WO2008076406A2 (en) 2006-12-14 2008-06-26 Ion Torrent Systems Incorporated Methods and apparatus for measuring analytes using large scale fet arrays
US8338166B2 (en) 2007-01-04 2012-12-25 Lawrence Livermore National Security, Llc Sorting, amplification, detection, and identification of nucleic acid subsequences in a complex mixture
JP2008167722A (ja) 2007-01-15 2008-07-24 Konica Minolta Medical & Graphic Inc 磁性支持体上での加熱による核酸単離方法
US7844658B2 (en) 2007-01-22 2010-11-30 Comcast Cable Holdings, Llc System and method for providing an application to a device
US20080176768A1 (en) 2007-01-23 2008-07-24 Honeywell Honeywell International Hydrogel microarray with embedded metal nanoparticles
WO2008093098A2 (en) 2007-02-02 2008-08-07 Illumina Cambridge Limited Methods for indexing samples and sequencing multiple nucleotide templates
US8003312B2 (en) 2007-02-16 2011-08-23 The Board Of Trustees Of The Leland Stanford Junior University Multiplex cellular assays using detectable cell barcodes
FI20075124A0 (fi) 2007-02-21 2007-02-21 Valtion Teknillinen Menetelmä ja testikitti nukleotidivariaatioiden toteamiseksi
US9029085B2 (en) 2007-03-07 2015-05-12 President And Fellows Of Harvard College Assays and other reactions involving droplets
US20080228268A1 (en) 2007-03-15 2008-09-18 Uluru, Inc. Method of Formation of Viscous, Shape Conforming Gels and Their Uses as Medical Prosthesis
US7943330B2 (en) 2007-03-23 2011-05-17 Academia Sinica Tailored glycoproteomic methods for the sequencing, mapping and identification of cellular glycoproteins
CN102014871A (zh) 2007-03-28 2011-04-13 哈佛大学 乳液及其形成技术
US9222936B2 (en) 2007-04-18 2015-12-29 Solulink, Inc. Methods and/or use of oligonucleotide conjugates for suppressing background due to cross-hybridization
WO2008134153A1 (en) 2007-04-23 2008-11-06 Advanced Liquid Logic, Inc. Bead-based multiplexed analytical methods and instrumentation
CN101293191B (zh) 2007-04-25 2011-11-09 中国科学院过程工程研究所 一种琼脂糖凝胶微球的制备方法
WO2008135512A2 (en) 2007-05-02 2008-11-13 Jerzy Paszkowski Dna amplification method
US20080295909A1 (en) 2007-05-24 2008-12-04 Locascio Laurie E Microfluidic Device for Passive Sorting and Storage of Liquid Plugs Using Capillary Force
CA2689356A1 (en) 2007-06-01 2008-12-11 454 Life Sciences Corporation System and meth0d for identification of individual samples from a multiplex mixture
WO2008148200A1 (en) 2007-06-05 2008-12-11 Eugenia Kumacheva Multiple continuous microfluidic reactors for the scaled up synthesis of gel or polymer particles
WO2009005680A1 (en) 2007-06-29 2009-01-08 President And Fellows Of Harvard College Methods and apparatus for manipulation of fluidic species
WO2009011808A1 (en) 2007-07-13 2009-01-22 President And Fellows Of Harvard College Droplet-based selection
US8454906B2 (en) 2007-07-24 2013-06-04 The Regents Of The University Of California Microfabricated droplet generator for single molecule/cell genetic analysis in engineered monodispersed emulsions
US20130084243A1 (en) 2010-01-27 2013-04-04 Liliane Goetsch Igf-1r specific antibodies useful in the detection and diagnosis of cellular proliferative disorders
AU2008286737A1 (en) 2007-08-15 2009-02-19 New York University Method, system and software arrangement for comparative analysis and phylogeny with whole-genome optical maps
US8268564B2 (en) 2007-09-26 2012-09-18 President And Fellows Of Harvard College Methods and applications for stitched DNA barcodes
WO2009048532A2 (en) 2007-10-05 2009-04-16 President And Fellows Of Harvard College Formation of particles for ultrasound application, drug release, and other uses, and microfluidic methods of preparation
US20090099040A1 (en) 2007-10-15 2009-04-16 Sigma Aldrich Company Degenerate oligonucleotides and their uses
US20100086914A1 (en) 2008-10-03 2010-04-08 Roche Molecular Systems, Inc. High resolution, high throughput hla genotyping by clonal sequencing
WO2009055732A1 (en) 2007-10-26 2009-04-30 Rosetta Inpharmatics Llc Cdna synthesis using non-random primers
US8334013B2 (en) 2007-11-02 2012-12-18 Stc.Unm Mesoporous metal oxide microspheres and method for forming same
WO2009061372A1 (en) 2007-11-02 2009-05-14 President And Fellows Of Harvard College Systems and methods for creating multi-phase entities, including particles and/or fluids
US8592150B2 (en) * 2007-12-05 2013-11-26 Complete Genomics, Inc. Methods and compositions for long fragment read sequencing
CN101918590B (zh) * 2007-12-10 2013-03-27 高晓莲 核酸测序
US7771944B2 (en) 2007-12-14 2010-08-10 The Board Of Trustees Of The University Of Illinois Methods for determining genetic haplotypes and DNA mapping
EP2235210B1 (en) 2007-12-21 2015-03-25 President and Fellows of Harvard College Methods for nucleic acid sequencing
ES2990227T3 (es) 2008-01-17 2024-11-29 Sequenom Inc Composiciones y procedimientos de análisis de ácido nucleico de secuencia única
KR20090081260A (ko) 2008-01-23 2009-07-28 삼성전자주식회사 마이크로 어레이 혼성화 검출 방법
US8501922B2 (en) 2008-02-07 2013-08-06 Pacific Biosciences Of California, Inc. CIS reactive oxygen quenchers integrated into linkers
JP5468271B2 (ja) 2008-02-08 2014-04-09 花王株式会社 微粒子分散液の製造方法
US20090203531A1 (en) 2008-02-12 2009-08-13 Nurith Kurn Method for Archiving and Clonal Expansion
AU2009224170B2 (en) 2008-03-11 2012-03-29 National Cancer Center Method for measuring chromosome, gene or specific nucleotide sequence copy numbers using SNP array
CN101241126A (zh) 2008-03-14 2008-08-13 东华大学 一种生物条形码探针的制备方法
US9011777B2 (en) 2008-03-21 2015-04-21 Lawrence Livermore National Security, Llc Monodisperse microdroplet generation and stopping without coalescence
US8961902B2 (en) 2008-04-23 2015-02-24 Bioscale, Inc. Method and apparatus for analyte processing
US9068181B2 (en) 2008-05-23 2015-06-30 The General Hospital Corporation Microfluidic droplet encapsulation
DE102008025656B4 (de) 2008-05-28 2016-07-28 Genxpro Gmbh Verfahren zur quantitativen Analyse von Nukleinsäuren, Marker dafür und deren Verwendung
GB0810051D0 (en) 2008-06-02 2008-07-09 Oxford Biodynamics Ltd Method of diagnosis
WO2009148598A1 (en) 2008-06-05 2009-12-10 President And Fellows Of Harvard College Polymersomes, colloidosomes, liposomes, and other species associated with fluidic droplets
US8198028B2 (en) 2008-07-02 2012-06-12 Illumina Cambridge Limited Using populations of beads for the fabrication of arrays on surfaces
WO2010009365A1 (en) 2008-07-18 2010-01-21 Raindance Technologies, Inc. Droplet libraries
WO2010009735A2 (en) 2008-07-23 2010-01-28 Dako Denmark A/S Combinatorial analysis and repair
US20100062494A1 (en) 2008-08-08 2010-03-11 President And Fellows Of Harvard College Enzymatic oligonucleotide pre-adenylation
US8383345B2 (en) 2008-09-12 2013-02-26 University Of Washington Sequence tag directed subassembly of short sequencing reads into long sequencing reads
WO2010033200A2 (en) 2008-09-19 2010-03-25 President And Fellows Of Harvard College Creation of libraries of droplets and related species
US8663920B2 (en) 2011-07-29 2014-03-04 Bio-Rad Laboratories, Inc. Library characterization by digital assay
US9764322B2 (en) 2008-09-23 2017-09-19 Bio-Rad Laboratories, Inc. System for generating droplets with pressure monitoring
US8709762B2 (en) * 2010-03-02 2014-04-29 Bio-Rad Laboratories, Inc. System for hot-start amplification via a multiple emulsion
US20120252015A1 (en) 2011-02-18 2012-10-04 Bio-Rad Laboratories Methods and compositions for detecting genetic material
US9089844B2 (en) 2010-11-01 2015-07-28 Bio-Rad Laboratories, Inc. System for forming emulsions
US9417190B2 (en) 2008-09-23 2016-08-16 Bio-Rad Laboratories, Inc. Calibrations and controls for droplet-based assays
WO2013016459A1 (en) 2011-07-25 2013-01-31 Bio-Rad Laboratories, Inc. Breakage of an emulsion containing nucleic acid
US9156010B2 (en) 2008-09-23 2015-10-13 Bio-Rad Laboratories, Inc. Droplet-based assay system
EP2889029A1 (en) 2008-09-25 2015-07-01 Cephalon, Inc. Liquid formulations of bendamustine
US8361299B2 (en) 2008-10-08 2013-01-29 Sage Science, Inc. Multichannel preparative electrophoresis system
US9080211B2 (en) 2008-10-24 2015-07-14 Epicentre Technologies Corporation Transposon end compositions and methods for modifying nucleic acids
CA2750054C (en) 2008-10-24 2018-05-29 Epicentre Technologies Corporation Transposon end compositions and methods for modifying nucleic acids
US20100113296A1 (en) 2008-11-05 2010-05-06 Joel Myerson Methods And Kits For Nucleic Acid Analysis
US8748103B2 (en) 2008-11-07 2014-06-10 Sequenta, Inc. Monitoring health and disease status using clonotype profiles
US8236503B2 (en) 2008-11-07 2012-08-07 Sequenta, Inc. Methods of monitoring conditions by sequence analysis
US20100203647A1 (en) 2008-11-21 2010-08-12 The Rockefeller University Chemical Reporters of Protein Acylation
CA2749570A1 (en) 2008-12-02 2010-06-10 Bio-Rad Laboratories, Inc. Chromatin structure detection
EP3587594B1 (en) 2008-12-19 2022-04-13 President and Fellows of Harvard College Particle-assisted nucleic acid sequencing
US10839940B2 (en) 2008-12-24 2020-11-17 New York University Method, computer-accessible medium and systems for score-driven whole-genome shotgun sequence assemble
KR101065807B1 (ko) 2009-01-23 2011-09-19 충남대학교산학협력단 액적 기반의 미세유체 칩을 이용한 마이크로 캡슐 제조방법
US9347092B2 (en) 2009-02-25 2016-05-24 Roche Molecular System, Inc. Solid support for high-throughput nucleic acid analysis
JP5457222B2 (ja) 2009-02-25 2014-04-02 エフ．ホフマン−ラ ロシュ アーゲー 小型化ハイスループット核酸分析
WO2010104597A2 (en) 2009-03-13 2010-09-16 President And Fellows Of Harvard College Scale-up of microfluidic devices
WO2010104604A1 (en) 2009-03-13 2010-09-16 President And Fellows Of Harvard College Method for the controlled creation of emulsions, including multiple emulsions
EP2230312A1 (en) 2009-03-19 2010-09-22 Helmholtz-Zentrum für Infektionsforschung GmbH Probe compound for detecting and isolating enzymes and means and methods using the same
WO2010111231A1 (en) 2009-03-23 2010-09-30 Raindance Technologies, Inc. Manipulation of microfluidic droplets
ES2903425T3 (es) 2009-03-30 2022-04-01 Illumina Inc Análisis de expresión génica en células individuales
KR101829182B1 (ko) * 2009-04-02 2018-03-29 플루이다임 코포레이션 표적 핵산의 바코딩을 위한 멀티 프라이머 증폭 방법
US9085798B2 (en) 2009-04-30 2015-07-21 Prognosys Biosciences, Inc. Nucleic acid constructs and methods of use
WO2010127304A2 (en) 2009-05-01 2010-11-04 Illumina, Inc. Sequencing methods
US9334531B2 (en) 2010-12-17 2016-05-10 Life Technologies Corporation Nucleic acid amplification
FR2945545B1 (fr) 2009-05-14 2011-08-05 Univ Aix Marseille Ii Methode de detection d'adn procaryote extrait d'un echantillon de selles
FR2945819B1 (fr) 2009-05-19 2011-06-17 Commissariat Energie Atomique Dispositif et procede d'isolement de cibles biologiques ou chimiques
US8574835B2 (en) 2009-05-29 2013-11-05 Life Technologies Corporation Scaffolded nucleic acid polymer particles and methods of making and using
DK2443236T3 (en) 2009-06-15 2015-08-17 Complete Genomics Inc Methods and compositions for sequencing by long fragment reading
US9524369B2 (en) 2009-06-15 2016-12-20 Complete Genomics, Inc. Processing and analysis of complex nucleic acid sequence data
EP2446278B1 (en) 2009-06-26 2021-11-17 President and Fellows of Harvard College Fluid injection
US20110028412A1 (en) 2009-08-03 2011-02-03 Cappellos, Inc. Herbal enhanced analgesic formulations
US20110033548A1 (en) * 2009-08-05 2011-02-10 E.I. Du Pont De Nemours And Company Degradable crosslinked aminated dextran microspheres and methods of use
US8298767B2 (en) 2009-08-20 2012-10-30 Population Genetics Technologies Ltd Compositions and methods for intramolecular nucleic acid rearrangement
WO2011027268A2 (en) 2009-09-01 2011-03-10 Koninklijke Philips Electronics N.V. Devices and methods for microarray selection
CA3021714C (en) * 2009-09-02 2021-03-09 Bio-Rad Laboratories, Inc. System for mixing fluids by coalescence of multiple emulsions
KR20120089661A (ko) 2009-09-02 2012-08-13 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 분출 및 다른 기술을 사용하여 생성되는 다중 에멀션
US9625454B2 (en) 2009-09-04 2017-04-18 The Research Foundation For The State University Of New York Rapid and continuous analyte processing in droplet microfluidic devices
GB0918564D0 (en) * 2009-10-22 2009-12-09 Plasticell Ltd Nested cell encapsulation
WO2011056546A1 (en) 2009-10-27 2011-05-12 President And Fellows Of Harvard College Droplet creation techniques
US10207240B2 (en) 2009-11-03 2019-02-19 Gen9, Inc. Methods and microfluidic devices for the manipulation of droplets in high fidelity polynucleotide assembly
CN107028886A (zh) 2009-11-04 2017-08-11 不列颠哥伦比亚大学 含有核酸的脂质粒子及相关的方法
CN102985552B (zh) 2009-11-25 2016-02-17 伯乐生命医学产品有限公司 用于检测遗传物质的方法和组合物
US9023769B2 (en) 2009-11-30 2015-05-05 Complete Genomics, Inc. cDNA library for nucleic acid sequencing
US8835358B2 (en) 2009-12-15 2014-09-16 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
EP2513333B1 (en) 2009-12-17 2013-10-02 Keygene N.V. Restriction enzyme based whole genome sequencing
WO2011079176A2 (en) 2009-12-23 2011-06-30 Raindance Technologies, Inc. Microfluidic systems and methods for reducing the exchange of molecules between droplets
EP4484577A3 (en) 2010-02-12 2025-03-26 Bio-Rad Laboratories, Inc. Digital analyte analysis
JP5901046B2 (ja) 2010-02-19 2016-04-06 国立大学法人 千葉大学 ＯＡＴＰ１Ｂ３ｍＲＮＡの新規な選択的スプライシングバリアント
US20110257889A1 (en) 2010-02-24 2011-10-20 Pacific Biosciences Of California, Inc. Sequence assembly and consensus sequence determination
AU2011219941C1 (en) 2010-02-24 2015-05-07 Arrowhead Pharmaceuticals, Inc. Compositions for targeted delivery of siRNA
HUE027972T2 (en) 2010-02-25 2016-11-28 Advanced Liquid Logic Inc A method for generating nucleic acid libraries
US8236574B2 (en) 2010-03-01 2012-08-07 Quanterix Corporation Ultra-sensitive detection of molecules or particles using beads or other capture objects
JP2013523651A (ja) 2010-03-24 2013-06-17 ノースイースタン ユニヴァーシティ 多重コンパートメントのマクロファージ送達
JP2013524171A (ja) 2010-03-25 2013-06-17 クァンタライフ・インコーポレーテッド 液滴ベースのアッセイのための液滴の発生
FR2958186A1 (fr) 2010-03-30 2011-10-07 Ecole Polytech Dispositif de formation de gouttes dans un circuit microfluide.
DK2556171T3 (en) 2010-04-05 2015-12-14 Prognosys Biosciences Inc Spatially CODED BIOLOGICAL ASSAYS
US9255291B2 (en) 2010-05-06 2016-02-09 Bioo Scientific Corporation Oligonucleotide ligation methods for improving data quality and throughput using massively parallel sequencing
US20110287947A1 (en) 2010-05-18 2011-11-24 University Of Southern California Tethered Conformation Capture
AU2011258165B2 (en) 2010-05-26 2016-11-17 Selecta Biosciences, Inc. Dose selection of adjuvanted synthetic nanocarriers
US20120000777A1 (en) 2010-06-04 2012-01-05 The Regents Of The University Of California Devices and methods for forming double emulsion droplet compositions and polymer particles
CN103119439A (zh) 2010-06-08 2013-05-22 纽亘技术公司 用于多重测序的方法和组合物
US8703493B2 (en) 2010-06-15 2014-04-22 Src, Inc. Location analysis using fire retardant-protected nucleic acid-labeled tags
US20120003657A1 (en) 2010-07-02 2012-01-05 Samuel Myllykangas Targeted sequencing library preparation by genomic dna circularization
US20120238738A1 (en) 2010-07-19 2012-09-20 New England Biolabs, Inc. Oligonucleotide Adapters: Compositions and Methods of Use
US20120034603A1 (en) 2010-08-06 2012-02-09 Tandem Diagnostics, Inc. Ligation-based detection of genetic variants
CN103202812B (zh) 2010-08-09 2015-10-28 南京大学 一种制备用于体内递送药理活性物质的蛋白纳米粒的方法
WO2012019765A1 (en) 2010-08-10 2012-02-16 European Molecular Biology Laboratory (Embl) Methods and systems for tracking samples and sample combinations
EP2616103A1 (en) 2010-09-16 2013-07-24 The University of North Carolina At Chapel Hill Asymmetric bifunctional silyl monomers and particles thereof as prodrugs and delivery vehicles for pharmaceutical, chemical and biological agents
WO2012038839A2 (en) 2010-09-21 2012-03-29 Population Genetics Technologies Ltd. Increasing confidence of allele calls with molecular counting
CN102409048B (zh) 2010-09-21 2013-10-23 深圳华大基因科技服务有限公司 一种基于高通量测序的dna标签文库构建方法
GB2499340B (en) 2010-10-04 2015-10-28 Genapsys Inc Methods for sequencing nucleic acids
US9999886B2 (en) 2010-10-07 2018-06-19 The Regents Of The University Of California Methods and systems for on demand droplet generation and impedance based detection
WO2012048341A1 (en) * 2010-10-08 2012-04-12 President And Fellows Of Harvard College High-throughput single cell barcoding
GB2498163B (en) 2010-10-08 2015-07-01 Harvard College High-throughput immune sequencing
CN102050953B (zh) 2010-10-18 2012-11-07 武汉理工大学 一种可还原降解超分子水凝胶的制备方法
US8753816B2 (en) 2010-10-26 2014-06-17 Illumina, Inc. Sequencing methods
US20130225623A1 (en) 2010-10-27 2013-08-29 Mount Sinai School Of Medicine Methods of Treating Psychiatric or Neurological Disorders with MGLUR Antagonists
AU2011323107B2 (en) * 2010-11-05 2015-09-10 Illumina, Inc. Linking sequence reads using paired code tags
US9074251B2 (en) 2011-02-10 2015-07-07 Illumina, Inc. Linking sequence reads using paired code tags
US8829171B2 (en) 2011-02-10 2014-09-09 Illumina, Inc. Linking sequence reads using paired code tags
EP2652155B1 (en) 2010-12-16 2016-11-16 Gigagen, Inc. Methods for massively parallel analysis of nucleic acids in single cells
US9163281B2 (en) 2010-12-23 2015-10-20 Good Start Genetics, Inc. Methods for maintaining the integrity and identification of a nucleic acid template in a multiplex sequencing reaction
CA2822439A1 (en) 2010-12-23 2012-06-28 Sequenom, Inc. Fetal genetic variation detection
WO2012100216A2 (en) 2011-01-20 2012-07-26 Knome, Inc. Methods and apparatus for assigning a meaningful numeric value to genomic variants, and searching and assessing same
GB201101429D0 (en) 2011-01-27 2011-03-16 Biocompatibles Uk Ltd Drug delivery system
US8765455B2 (en) 2011-01-27 2014-07-01 Lawrence Livermore National Security, Llc Chip-based droplet sorting
AU2012211081B2 (en) 2011-01-28 2016-02-04 Illumina, Inc. Oligonucleotide replacement for di-tagged and directional libraries
JP6069224B2 (ja) 2011-01-31 2017-02-01 アプライズ バイオ， インコーポレイテッド 細胞において複数のエピトープを同定する方法
EP2670894B1 (en) * 2011-02-02 2017-11-29 University Of Washington Through Its Center For Commercialization Massively parallel continguity mapping
EP3940084A1 (en) 2011-02-09 2022-01-19 Bio-Rad Laboratories, Inc. Analysis of nucleic acids
EP3859011A1 (en) 2011-02-11 2021-08-04 Bio-Rad Laboratories, Inc. Methods for forming mixed droplets
US9266104B2 (en) 2011-02-11 2016-02-23 Raindance Technologies, Inc. Thermocycling device for nucleic acid amplification and methods of use
EP2675819B1 (en) 2011-02-18 2020-04-08 Bio-Rad Laboratories, Inc. Compositions and methods for molecular labeling
WO2012116331A2 (en) 2011-02-25 2012-08-30 Illumina, Inc. Methods and systems for haplotype determination
WO2012116250A1 (en) 2011-02-25 2012-08-30 University Of Massachusetts Medical School Monomers and polymers for functional polycarbonates and poly (ester-carbonates) and peg-co-polycarbonate hydrogels
WO2012121973A1 (en) 2011-03-04 2012-09-13 Life Technologies Corporation Compounds and methods for conjugation of biomolecules
US20120233202A1 (en) 2011-03-09 2012-09-13 Annai Systems, Inc. Biological data networks and methods therefor
JP2014509865A (ja) 2011-03-18 2014-04-24 バイオ−ラッド・ラボラトリーズ・インコーポレーテッド シグナルの組合せ使用による多重化デジタルアッセイ
US9260753B2 (en) 2011-03-24 2016-02-16 President And Fellows Of Harvard College Single cell nucleic acid detection and analysis
GB2489714B (en) * 2011-04-05 2013-11-06 Tracesa Ltd Fluid Identification Method
WO2012150317A1 (en) 2011-05-05 2012-11-08 Institut National De La Sante Et De La Recherche Medicale (Inserm) Linear dna amplification
GB201106254D0 (en) 2011-04-13 2011-05-25 Frisen Jonas Method and product
CN107368705B (zh) 2011-04-14 2021-07-13 完整基因有限公司 分析生物体的基因组dna的方法和计算机系统
EP3789498B1 (en) 2011-04-25 2025-08-13 Bio-Rad Laboratories, Inc. Methods for nucleic acid analysis
WO2012149438A1 (en) 2011-04-28 2012-11-01 Life Technologies Corporation Methods and compositions for multiplex pcr
PL3418380T3 (pl) 2011-04-28 2022-09-19 The Board Of Trustees Of The Leland Stanford Junior University Identyfikacja polinukleotydów powiązanych z próbką
AU2012257416B8 (en) 2011-05-16 2017-10-19 Tagworks Pharmaceuticals B.V. Bio-orthogonal drug activation
JP6100685B2 (ja) 2011-05-16 2017-03-22 地方独立行政法人 大阪府立病院機構 血中ｄｎａの定量的検出による悪性新生物の病勢の進行を評価する方法
KR20140034242A (ko) 2011-05-23 2014-03-19 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 다중 에멀젼을 포함하는 에멀젼의 제어
US9005935B2 (en) 2011-05-23 2015-04-14 Agilent Technologies, Inc. Methods and compositions for DNA fragmentation and tagging by transposases
WO2012166425A2 (en) 2011-05-27 2012-12-06 President And Fellows Of Harvard College Methods of amplifying whole genome of a single cell
US8841071B2 (en) 2011-06-02 2014-09-23 Raindance Technologies, Inc. Sample multiplexing
EP3709018A1 (en) 2011-06-02 2020-09-16 Bio-Rad Laboratories, Inc. Microfluidic apparatus for identifying components of a chemical reaction
US9150916B2 (en) 2011-06-24 2015-10-06 Beat Christen Compositions and methods for identifying the essential genome of an organism
US8927218B2 (en) 2011-06-27 2015-01-06 Flir Systems, Inc. Methods and compositions for segregating target nucleic acid from mixed nucleic acid samples
WO2013006824A2 (en) 2011-07-07 2013-01-10 Life Technologies Corporation Polymer particles, nucleic acid polymer particles and methods of making and using the same
US20130017978A1 (en) 2011-07-11 2013-01-17 Finnzymes Oy Methods and transposon nucleic acids for generating a dna library
US9605304B2 (en) 2011-07-20 2017-03-28 The Hong Kong Polytechnic University Ultra-stable oligonucleotide-gold and-silver nanoparticle conjugates and method of their preparation
US8658430B2 (en) 2011-07-20 2014-02-25 Raindance Technologies, Inc. Manipulating droplet size
US9012373B2 (en) 2011-08-04 2015-04-21 Sage Science, Inc. Systems and methods for processing fluids
US10364464B2 (en) 2011-08-08 2019-07-30 The Broad Institute, Inc. Compositions and methods for co-amplifying subsequences of a nucleic acid fragment sequence
US20120048340A1 (en) * 2011-08-09 2012-03-01 General Electric Company Solar panel tracking system and associated tracking sensor
US8846883B2 (en) 2011-08-16 2014-09-30 University Of Southhampton Oligonucleotide ligation
WO2013035114A1 (en) 2011-09-08 2013-03-14 Decode Genetics Ehf Tp53 genetic variants predictive of cancer
US9249460B2 (en) 2011-09-09 2016-02-02 The Board Of Trustees Of The Leland Stanford Junior University Methods for obtaining a sequence
EP2768607B1 (en) 2011-09-26 2021-08-18 Thermo Fisher Scientific GENEART GmbH Multiwell plate for high efficiency, small volume nucleic acid synthesis
EP2760578B1 (en) 2011-09-28 2020-08-26 President and Fellows of Harvard College Systems and methods for droplet production and/or fluidic manipulation
US9514272B2 (en) 2011-10-12 2016-12-06 Complete Genomics, Inc. Identification of DNA fragments and structural variations
US9469874B2 (en) 2011-10-18 2016-10-18 The Regents Of The University Of California Long-range barcode labeling-sequencing
US20130109576A1 (en) 2011-10-28 2013-05-02 Anthony P. Shuber Methods for detecting mutations
WO2013066416A1 (en) 2011-11-04 2013-05-10 Intel Corporation Signaling for configuration of downlink coordinated multipoint communications
JP6083713B2 (ja) 2011-11-16 2017-02-22 インターナショナル・ビジネス・マシーンズ・コーポレーションＩｎｔｅｒｎａｔｉｏｎａｌ Ｂｕｓｉｎｅｓｓ Ｍａｃｈｉｎｅｓ Ｃｏｒｐｏｒａｔｉｏｎ 変形可能バルブを備えたマイクロ流体デバイス
US10689643B2 (en) 2011-11-22 2020-06-23 Active Motif, Inc. Targeted transposition for use in epigenetic studies
US9938524B2 (en) 2011-11-22 2018-04-10 Active Motif, Inc. Multiplex isolation of protein-associated nucleic acids
CN107345204B (zh) 2011-12-03 2021-02-12 Emd密理博公司 用于微流体细胞培养的微型培育系统和方法
ES2953308T3 (es) 2011-12-22 2023-11-10 Harvard College Composiciones y métodos para la detección de analitos
WO2013096643A1 (en) 2011-12-23 2013-06-27 Gigagen Methods and apparatuses for droplet mixing
JP6170074B2 (ja) 2012-02-09 2017-07-26 ライフ テクノロジーズ コーポレーション 親水性ポリマー粒子およびその製造方法
CN104271768A (zh) 2012-02-14 2015-01-07 约翰霍普金斯大学 miRNA分析方法
CA2864287C (en) 2012-02-15 2021-01-12 Wisconsin Alumni Research Foundation Dithioamine reducing agents
US10011871B2 (en) 2012-02-17 2018-07-03 Fred Hutchinson Cancer Research Center Compositions and methods for accurately identifying mutations
US10202628B2 (en) 2012-02-17 2019-02-12 President And Fellows Of Harvard College Assembly of nucleic acid sequences in emulsions
EP2817418B1 (en) 2012-02-24 2017-10-11 Raindance Technologies, Inc. Labeling and sample preparation for sequencing
EP3305918B1 (en) 2012-03-05 2020-06-03 President and Fellows of Harvard College Methods for epigenetic sequencing
NO2694769T3 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) 2012-03-06 2018-03-03
US9552458B2 (en) 2012-03-16 2017-01-24 The Research Institute At Nationwide Children's Hospital Comprehensive analysis pipeline for discovery of human genetic variation
US20150125904A1 (en) 2012-03-30 2015-05-07 Massachusetts Institute Of Technology Probe incorporation mediated by enzymes
CN102622634B (zh) * 2012-03-31 2014-04-23 中国农业科学院果树研究所 一种苹果种质资源条形码标识的制备方法
EP2647426A1 (en) 2012-04-03 2013-10-09 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Replication of distributed nucleic acid molecules with preservation of their relative distribution through hybridization-based binding
US8209130B1 (en) 2012-04-04 2012-06-26 Good Start Genetics, Inc. Sequence assembly
JP2015519886A (ja) 2012-04-16 2015-07-16 バイオロジカル ダイナミクス，インク． 核酸サンプル調製
US20130296173A1 (en) 2012-04-23 2013-11-07 Complete Genomics, Inc. Pre-anchor wash
EP4148142B1 (en) 2012-05-21 2025-08-13 The Scripps Research Institute Methods of sample preparation
JP6558830B2 (ja) 2012-06-15 2019-08-14 ボード・オブ・リージエンツ，ザ・ユニバーシテイ・オブ・テキサス・システム 複数転写産物のハイスループットシークエンシング
SG11201408128WA (en) 2012-06-15 2015-01-29 Adaptive Biotechnologies Corp Uniquely tagged rearranged adaptive immune receptor genes in a complex gene set
WO2014018460A1 (en) 2012-07-24 2014-01-30 Sequenta, Inc. Single cell analysis using sequence tags
EP3578669B1 (en) 2012-08-08 2024-07-10 F. Hoffmann-La Roche AG Increasing dynamic range for identifying multiple epitopes in cells
JP6514105B2 (ja) 2012-08-13 2019-05-15 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 生物学的成分を検出するための方法およびシステム
US10273541B2 (en) 2012-08-14 2019-04-30 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10323279B2 (en) 2012-08-14 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10752949B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9701998B2 (en) 2012-12-14 2017-07-11 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9951386B2 (en) 2014-06-26 2018-04-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
US20150005199A1 (en) 2012-08-14 2015-01-01 10X Technologies, Inc. Compositions and methods for sample processing
US10221442B2 (en) 2012-08-14 2019-03-05 10X Genomics, Inc. Compositions and methods for sample processing
US20140378345A1 (en) 2012-08-14 2014-12-25 10X Technologies, Inc. Compositions and methods for sample processing
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
US10400280B2 (en) 2012-08-14 2019-09-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
CA2881685C (en) 2012-08-14 2023-12-05 10X Genomics, Inc. Microcapsule compositions and methods
US20150005200A1 (en) 2012-08-14 2015-01-01 10X Technologies, Inc. Compositions and methods for sample processing
US20220098659A1 (en) 2012-08-14 2022-03-31 10X Genomics, Inc. Methods and systems for processing polynucleotides
US20140378349A1 (en) 2012-08-14 2014-12-25 10X Technologies, Inc. Compositions and methods for sample processing
US20140378322A1 (en) 2012-08-14 2014-12-25 10X Technologies, Inc. Compositions and methods for sample processing
EP2898096B1 (en) 2012-09-21 2024-02-14 The Broad Institute, Inc. Methods for labeling of rnas
US9644199B2 (en) 2012-10-01 2017-05-09 Agilent Technologies, Inc. Immobilized transposase complexes for DNA fragmentation and tagging
GB201217772D0 (en) 2012-10-04 2012-11-14 Base4 Innovation Ltd Sequencing method
FR2996544B1 (fr) 2012-10-08 2015-03-13 Ecole Polytech Circuit microfluidique permettant la mise en contact de gouttes de plusieurs fluides, et procede microfluidique correspondant.
FR2996545B1 (fr) 2012-10-08 2016-03-25 Ecole Polytech Procede microfluidique de traitement et d'analyse d'une solution contenant un materiel biologique, et circuit microfluidique correspondant.
WO2014062717A1 (en) 2012-10-15 2014-04-24 Life Technologies Corporation Compositions, methods, systems and kits for target nucleic acid enrichment
USRE50065E1 (en) 2012-10-17 2024-07-30 10X Genomics Sweden Ab Methods and product for optimising localised or spatial detection of gene expression in a tissue sample
CN104903466B (zh) 2012-11-05 2016-11-23 鲁比康基因组学公司 条形编码核酸
EP2917366B1 (en) 2012-11-06 2017-08-02 Oxford Nanopore Technologies Limited Quadruplex method
EP2917368A1 (en) 2012-11-07 2015-09-16 Good Start Genetics, Inc. Methods and systems for identifying contamination in samples
HK1217112A1 (zh) 2012-12-03 2016-12-23 以琳生物药物有限公司 单链多核苷酸扩增方法
WO2014093701A1 (en) 2012-12-12 2014-06-19 The Broad Institute, Inc. Functional genomics using crispr-cas systems, compositions, methods, knock out libraries and applications thereof
CN105121648B (zh) 2012-12-12 2021-05-07 布罗德研究所有限公司 用于序列操纵的系统、方法和优化的指导组合物的工程化
US10533221B2 (en) 2012-12-14 2020-01-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
CA2894694C (en) 2012-12-14 2023-04-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
EP2749653A1 (en) 2012-12-28 2014-07-02 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Molecular coding for analysis of composition of macromolecules and molecular complexes
EP2752664A1 (en) 2013-01-07 2014-07-09 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Label-free method for the detection of analytes
US9683230B2 (en) 2013-01-09 2017-06-20 Illumina Cambridge Limited Sample preparation on a solid support
GB202020510D0 (en) 2013-01-17 2021-02-03 Edico Genome Corp Bioinformatics systems, apparatuses, and methods executed on an integrated circuit processing platform
KR101984699B1 (ko) 2013-01-24 2019-05-31 삼성전자주식회사 핵산 분석용 미세 유체 시스템
HK1218151A1 (zh) 2013-01-25 2017-02-03 Bio-Rad Laboratories, Inc. 实施液滴膨胀的系统和方法
US10381106B2 (en) 2013-01-28 2019-08-13 Hasso-Plattner-Institut Fuer Softwaresystemtechnik Gmbh Efficient genomic read alignment in an in-memory database
US11110458B2 (en) 2013-02-01 2021-09-07 Bio-Rad Laboratories, Inc. System for detection of spaced droplets
JP2016511243A (ja) 2013-02-08 2016-04-14 テンエックス・ジェノミクス・インコーポレイテッド ポリヌクレオチドバーコード生成
WO2014138154A1 (en) 2013-03-06 2014-09-12 President And Fellows Of Harvard College Devices and methods for forming relatively monodisperse droplets
AU2014225372B2 (en) 2013-03-08 2019-10-24 Bio-Rad Laboratories, Inc. Compositions, methods and systems for polymerase chain reaction assays
AU2013382098B2 (en) 2013-03-13 2019-02-07 Illumina, Inc. Methods and compositions for nucleic acid sequencing
US10612088B2 (en) 2013-03-14 2020-04-07 The Broad Institute, Inc. Massively multiplexed RNA sequencing
US9273349B2 (en) 2013-03-14 2016-03-01 Affymetrix, Inc. Detection of nucleic acids
WO2014145078A1 (en) 2013-03-15 2014-09-18 Verinata Health, Inc. Generating cell-free dna libraries directly from blood
JP6587544B2 (ja) 2013-03-15 2019-10-09 シグニス バイオテク エセ．エレ．ウ．Ｓｙｇｎｉｓ Ｂｉｏｔｅｃｈ Ｓ．Ｌ．Ｕ． 熱安定なＴｔｈＰｒｉｍＰｏｌを用いた増幅及び配列決定方法
US20140274729A1 (en) 2013-03-15 2014-09-18 Nugen Technologies, Inc. Methods, compositions and kits for generation of stranded rna or dna libraries
WO2014145760A1 (en) 2013-03-15 2014-09-18 Bio-Rad Laboratories, Inc. Droplet generator with collection tube
GB2525568B (en) 2013-03-15 2020-10-14 Abvitro Llc Single cell barcoding for antibody discovery
EP2972366B1 (en) 2013-03-15 2020-06-17 Prognosys Biosciences, Inc. Methods for detecting peptide/mhc/tcr binding
US9328382B2 (en) 2013-03-15 2016-05-03 Complete Genomics, Inc. Multiple tagging of individual long DNA fragments
EP4566715A3 (en) 2013-04-02 2025-08-27 Bio-Rad Laboratories, Inc. Systems and methods for handling microfluidic droplets
WO2014182835A1 (en) 2013-05-09 2014-11-13 Bio-Rad Laboratories, Inc. Magnetic immuno digital pcr assay
US10059989B2 (en) 2013-05-23 2018-08-28 The Board Of Trustees Of The Leland Stanford Junior University Transposition of native chromatin for personal epigenomics
JP6600302B2 (ja) 2013-06-12 2019-10-30 ザ ジェネラル ホスピタル コーポレイション 標的分子のマルチプレックス検出のための方法、キット、およびシステム、ならびにそれらの使用
WO2014201273A1 (en) 2013-06-12 2014-12-18 The Broad Institute, Inc. High-throughput rna-seq
CN107995927B (zh) 2013-06-17 2021-07-30 布罗德研究所有限公司 用于肝靶向和治疗的crispr-cas系统、载体和组合物的递送与用途
US20160208323A1 (en) 2013-06-21 2016-07-21 The Broad Institute, Inc. Methods for Shearing and Tagging DNA for Chromatin Immunoprecipitation and Sequencing
WO2014210225A1 (en) 2013-06-25 2014-12-31 Prognosys Biosciences, Inc. Methods and systems for determining spatial patterns of biological targets in a sample
EP3467160A1 (en) 2013-06-27 2019-04-10 10X Genomics, Inc. Compositions and methods for sample processing
US9840718B2 (en) 2013-07-12 2017-12-12 University Of South Alabama Minimal piggyBac vectors for genome integration
CN103394410B (zh) 2013-07-25 2016-04-20 博奥生物集团有限公司 一种位置可调节的智能磁力架
GB2516684A (en) 2013-07-30 2015-02-04 Sphere Fluidics Ltd Microfluidic devices and systems
KR102758333B1 (ko) 2013-08-28 2025-01-23 벡톤 디킨슨 앤드 컴퍼니 대량의 동시 단일 세포 분석
US10395758B2 (en) 2013-08-30 2019-08-27 10X Genomics, Inc. Sequencing methods
US9381217B2 (en) 2013-09-09 2016-07-05 Georgia Tech Research Corporation Microgels for encapsulation of cells and other biologic agents
WO2015048173A2 (en) 2013-09-24 2015-04-02 The Regents Of The University Of California Encapsulated sensors and sensing systems for bioassays and diagnostics and methods for making and using them
GB201317301D0 (en) 2013-09-30 2013-11-13 Linnarsson Sten Method for capturing and encoding nucleic acid from a plurality of single cells
CA2926934A1 (en) 2013-10-09 2015-04-16 Stc.Unm Synthetic long read dna sequencing
US9937495B2 (en) 2013-10-28 2018-04-10 Massachusetts Institute Of Technology Hydrogel microstructures with immiscible fluid isolation for small reaction volumes
US9824068B2 (en) 2013-12-16 2017-11-21 10X Genomics, Inc. Methods and apparatus for sorting data
US20140315755A1 (en) 2013-12-26 2014-10-23 Tao Chen Genome-wide Antisense Oligonucleotide and RNAi
PT3089822T (pt) 2013-12-30 2022-05-06 Atreca Inc Análise de ácidos nucleicos associados com células isoladas usando códigos de barras de ácido nucleico
KR101464100B1 (ko) 2014-01-29 2014-11-21 성균관대학교산학협력단 생체 적용 가능한 리포솜-핵산형광 나노 융합체, 이의 응용, 및 이의 제조방법
CN106103713B (zh) 2014-02-03 2021-05-28 赛默飞世尔科技波罗的海封闭股份公司 用于经控制dna片段化的方法
CN105531408B (zh) 2014-02-13 2019-09-10 生物辐射实验室股份有限公司 染色体构象划分产物捕获
US20160376663A1 (en) 2014-02-27 2016-12-29 Igenomx International Genomics Corporation Methods for analysis of somatic mobile elements, and uses thereof
KR102596508B1 (ko) 2014-04-10 2023-10-30 10엑스 제노믹스, 인크. 시약을 캡슐화 및 구획화하기 위한 유체 디바이스, 시스템, 및 방법, 및 이의 응용
TWI682997B (zh) 2014-04-15 2020-01-21 美商伊路米納有限公司 用於改善插入序列傾向及增加ｄｎａ輸入耐受度之經修飾的轉位酶
EP3134536B1 (en) 2014-04-21 2019-12-18 President and Fellows of Harvard College Systems and methods for barcoding nucleic acids
US20150298091A1 (en) 2014-04-21 2015-10-22 President And Fellows Of Harvard College Systems and methods for barcoding nucleic acids
US10975371B2 (en) 2014-04-29 2021-04-13 Illumina, Inc. Nucleic acid sequence analysis from single cells
US10526601B2 (en) 2014-05-23 2020-01-07 Digenomix Corporation Haploidome determination by digitized transposons
EP3152232B1 (en) 2014-06-06 2019-11-13 Herlev Hospital Determining antigen recognition through barcoding of mhc multimers
WO2015191877A1 (en) 2014-06-11 2015-12-17 Life Technologies Corporation Systems and methods for substrate enrichment
CA2950043C (en) 2014-06-11 2022-08-02 Samplix S.A.R.L. Nucleotide sequence exclusion enrichment by droplet sorting (needls)
EP4270006A3 (en) 2014-06-13 2024-01-10 Immudex ApS General detection and isolation of specific cells by binding of labeled molecules
US10480021B2 (en) 2014-06-23 2019-11-19 Yale University Methods for closed chromatin mapping and DNA methylation analysis for single cells
US11155809B2 (en) 2014-06-24 2021-10-26 Bio-Rad Laboratories, Inc. Digital PCR barcoding
CN106574298A (zh) 2014-06-26 2017-04-19 10X基因组学有限公司 用于样品分析的方法和组合物
US10017759B2 (en) 2014-06-26 2018-07-10 Illumina, Inc. Library preparation of tagged nucleic acid
CN106795553B (zh) 2014-06-26 2021-06-04 10X基因组学有限公司 分析来自单个细胞或细胞群体的核酸的方法
JP2017522866A (ja) 2014-06-26 2017-08-17 １０エックス ジェノミクス， インコーポレイテッド 核酸配列の分析
US10839939B2 (en) 2014-06-26 2020-11-17 10X Genomics, Inc. Processes and systems for nucleic acid sequence assembly
WO2016011414A1 (en) 2014-07-18 2016-01-21 Illumina, Inc. Non-invasive prenatal diagnosis of fetal genetic condition using cellular dna and cell free dna
US20160024558A1 (en) 2014-07-23 2016-01-28 10X Genomics, Inc. Nucleic acid binding proteins and uses thereof
CN105335116B (zh) 2014-07-30 2018-11-09 联想(北京)有限公司 一种显示控制方法及电子设备
EP3186418A2 (en) 2014-08-26 2017-07-05 Nugen Technologies, Inc. Compositions and methods for targeted nucleic acid sequence enrichment and high efficiency library generation
JP2017532024A (ja) 2014-09-09 2017-11-02 ザ・ブロード・インスティテュート・インコーポレイテッド コンポジット単一細胞核酸分析のための液滴ベースの方法および機器
AU2015318011B2 (en) 2014-09-15 2020-07-23 Abvitro Llc High-throughput nucleotide library sequencing
SG11201703139VA (en) 2014-10-17 2017-07-28 Illumina Cambridge Ltd Contiguity preserving transposition
AU2015339148B2 (en) 2014-10-29 2022-03-10 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequencing
US9975122B2 (en) 2014-11-05 2018-05-22 10X Genomics, Inc. Instrument systems for integrated sample processing
EP3234602B1 (en) 2014-12-19 2021-08-25 F. Hoffmann-La Roche AG Methods for identifying multiple epitopes in selected sub-populations of cells
BR112017014902A2 (pt) 2015-01-12 2018-03-13 10X Genomics Inc processos e sistemas para a preparação de bibliotecas de sequenciamento de ácido nucleico e bibliotecas preparadas usando os mesmos
MX2017008916A (es) 2015-01-13 2017-10-19 10X Genomics Inc Sistemas y metodos para visualizar informacion de fases y variaciones estructurales.
WO2016126882A1 (en) 2015-02-04 2016-08-11 Twist Bioscience Corporation Methods and devices for de novo oligonucleic acid assembly
JP2018508198A (ja) 2015-02-04 2018-03-29 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 別個の実体におけるバーコード付加による核酸のシーケンシング
US10854315B2 (en) 2015-02-09 2020-12-01 10X Genomics, Inc. Systems and methods for determining structural variation and phasing using variant call data
CN107406890B (zh) 2015-02-10 2023-07-18 亿明达股份有限公司 用于分析细胞组分的方法和组合物
US11274343B2 (en) 2015-02-24 2022-03-15 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequence coverage
EP3262407B1 (en) 2015-02-24 2023-08-30 10X Genomics, Inc. Partition processing methods and systems
EP3262192B1 (en) 2015-02-27 2020-09-16 Becton, Dickinson and Company Spatially addressable molecular barcoding
CN107614700A (zh) 2015-03-11 2018-01-19 布罗德研究所有限公司 基因型和表型偶联
WO2016149661A1 (en) 2015-03-18 2016-09-22 The Broad Institute, Inc. Massively parallel on-chip coalescence of microemulsions
US20160289769A1 (en) 2015-03-30 2016-10-06 Verily Life Sciences Llc Methods for Combining Single Cell Profiling with Combinatorial Nanoparticle Conjugate Library Screening and In Vivo Diagnostic System
ES2994053T3 (en) 2015-04-10 2025-01-17 10X Genomics Sweden Ab Spatially distinguished, multiplex nucleic acid analysis of biological specimens
EP3283629A4 (en) 2015-04-17 2018-08-29 President and Fellows of Harvard College Barcoding systems and methods for gene sequencing and other applications
CN107250447B (zh) 2015-04-20 2020-05-05 深圳华大生命科学研究院 一种长片段dna文库构建方法
US10632465B2 (en) 2015-04-22 2020-04-28 Stilla Technologies Contact-less priming method for loading a solution in a microfluidic device and associated system
US20160314242A1 (en) 2015-04-23 2016-10-27 10X Genomics, Inc. Sample indexing methods and compositions for sequencing applications
US20170016041A1 (en) 2015-05-18 2017-01-19 10X Genomics, Inc. Stabilized reducing agents and methods using same
EP3298164A2 (en) 2015-05-18 2018-03-28 10X Genomics, Inc. Mobile solid phase compositions for use in biochemical reactions and analyses
WO2016187717A1 (en) 2015-05-26 2016-12-01 Exerkine Corporation Exosomes useful for genome editing
WO2016191380A1 (en) 2015-05-26 2016-12-01 Pacific Biosciences Of California, Inc. De novo diploid genome assembly and haplotype sequence reconstruction
WO2016191618A1 (en) 2015-05-27 2016-12-01 Jianbiao Zheng Methods of inserting molecular barcodes
JP2018518992A (ja) 2015-06-24 2018-07-19 オックスフォード バイオダイナミックス リミテッド 染色体相互作用の部位を用いた検出法
EP3325665B1 (en) 2015-07-17 2020-09-02 President and Fellows of Harvard College Methods of amplifying nucleic acid sequences
SMT202000037T1 (it) 2015-08-12 2020-03-13 Cemm Forschungszentrum Fuer Molekulare Medizin Gmbh Metodi per studiare gli acidi nucleici
WO2017034970A1 (en) 2015-08-21 2017-03-02 The General Hospital Corporation Combinatorial single molecule analysis of chromatin
WO2017053905A1 (en) 2015-09-24 2017-03-30 Abvitro Llc Affinity-oligonucleotide conjugates and uses thereof
EP3362032A4 (en) 2015-10-13 2019-05-01 President and Fellows of Harvard College SYSTEMS AND METHOD FOR THE PRODUCTION AND USE OF GEL MICROBALLETS
EP3366818B1 (en) 2015-10-19 2021-04-14 Zhejiang Annoroad Bio-Technology Co., Ltd. Method for constructing high-resolution single cell hi-c library with a lot of information
EP3365454A1 (en) 2015-10-20 2018-08-29 10X Genomics, Inc. Methods and systems for high throughput single cell genetic manipulation
US11092607B2 (en) 2015-10-28 2021-08-17 The Board Institute, Inc. Multiplex analysis of single cell constituents
WO2017075294A1 (en) 2015-10-28 2017-05-04 The Board Institute Inc. Assays for massively combinatorial perturbation profiling and cellular circuit reconstruction
CN108473984A (zh) 2015-11-04 2018-08-31 阿特雷卡公司 用于分析与单个细胞相关联的核酸的核酸条形码的组合组
WO2017087910A1 (en) 2015-11-19 2017-05-26 10X Genomics, Inc. Transformable tagging compositions, methods, and processes incorporating same
EP3384048B1 (en) 2015-12-04 2021-03-24 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
EP3397764A4 (en) 2015-12-30 2019-05-22 Bio-Rad Laboratories, Inc. DIGITAL PROTEIN QUANTIFICATION
WO2017139690A1 (en) 2016-02-11 2017-08-17 10X Genomics, Inc. Cell population analysis using single nucleotide polymorphisms from single cell transcriptomes
EP3414341A4 (en) 2016-02-11 2019-10-09 10X Genomics, Inc. SYSTEMS, METHODS AND MEDIA FOR DE-NOVO ARRANGEMENT OF ALL GENERIC SEQUENCE DATA
EP3423598B1 (en) 2016-03-01 2024-05-01 Universal Sequencing Technology Corporation Methods and kits for tracking nucleic acid target origin for nucleic acid sequencing
CN109072206A (zh) 2016-03-10 2018-12-21 斯坦福大学托管董事会 转座酶介导的对可接近基因组的成像
CA3020542A1 (en) 2016-04-11 2017-10-19 Board Of Regents, The University Of Texas System Methods and compositions for detecting single t cell receptor affinity and sequence
JP2019514002A (ja) 2016-04-15 2019-05-30 プレジデント アンド フェローズ オブ ハーバード カレッジ 液滴および／または他の実体の収集のためのシステムおよび方法
JP2019515674A (ja) 2016-04-19 2019-06-13 プレジデント アンド フェローズ オブ ハーバード カレッジ 遺伝的解析および他の適用のための固定化ベースのシステムおよび方法
WO2017197343A2 (en) 2016-05-12 2017-11-16 10X Genomics, Inc. Microfluidic on-chip filters
WO2017197338A1 (en) 2016-05-13 2017-11-16 10X Genomics, Inc. Microfluidic systems and methods of use
CA3033506A1 (en) 2016-08-10 2018-02-15 President And Fellows Of Harvard College Methods of de novo assembly of barcoded genomic dna fragments
WO2018039338A1 (en) 2016-08-23 2018-03-01 10X Genomics, Inc. Microfluidic surface-mediated emulsion stability control
WO2018044831A1 (en) 2016-08-30 2018-03-08 Integrated Dna Technologies, Inc. Cleavable hairpin primers
CN118853848A (zh) 2016-08-31 2024-10-29 哈佛学院董事及会员团体 将生物分子的检测组合到使用荧光原位测序的单个试验的方法
AU2016421196A1 (en) 2016-08-31 2019-03-21 President And Fellows Of Harvard College Methods of whole genome digital amplification
US20180080021A1 (en) 2016-09-17 2018-03-22 The Board Of Trustees Of The Leland Stanford Junior University Simultaneous sequencing of rna and dna from the same sample
ES2961743T3 (es) 2016-09-26 2024-03-13 Becton Dickinson Co Medición de la expresión de proteínas utilizando reactivos con secuencias de oligonucleótidos con código de barras
CN109983126A (zh) 2016-10-19 2019-07-05 10X基因组学有限公司 用于条形码化单个细胞或细胞群的核酸分子的方法和系统
GB201619458D0 (en) 2016-11-17 2017-01-04 Spatial Transcriptomics Ab Method for spatial tagging and analysing nucleic acids in a biological specimen
CN109996892B (zh) 2016-12-07 2023-08-29 深圳华大智造科技股份有限公司 单细胞测序文库的构建方法及其应用
US11021738B2 (en) 2016-12-19 2021-06-01 Bio-Rad Laboratories, Inc. Droplet tagging contiguity preserved tagmented DNA
AU2017382905A1 (en) 2016-12-21 2019-07-04 The Regents Of The University Of California Single cell genomic sequencing using hydrogel based droplets
US10815525B2 (en) 2016-12-22 2020-10-27 10X Genomics, Inc. Methods and systems for processing polynucleotides
US20190177800A1 (en) 2017-12-08 2019-06-13 10X Genomics, Inc. Methods and compositions for labeling cells
US10550429B2 (en) 2016-12-22 2020-02-04 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10011872B1 (en) 2016-12-22 2018-07-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
KR102747205B1 (ko) 2016-12-29 2024-12-31 일루미나, 인코포레이티드 세포 구획 내의 생체분자에 대한 직교 접근 및 그의 태그부착을 위한 분석 시스템
EP3565907B1 (en) 2017-01-06 2022-05-04 Editas Medicine, Inc. Methods of assessing nuclease cleavage
US20210087610A9 (en) 2017-01-12 2021-03-25 Massachusetts Institute Of Technology Methods for analyzing t cell receptors and b cell receptors
WO2018140966A1 (en) 2017-01-30 2018-08-02 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
EP4324931A3 (en) 2017-02-02 2024-05-29 New York Genome Center, Inc. Methods and compositions for identifying or quantifying targets in a biological sample
US10995333B2 (en) 2017-02-06 2021-05-04 10X Genomics, Inc. Systems and methods for nucleic acid preparation
US10347365B2 (en) 2017-02-08 2019-07-09 10X Genomics, Inc. Systems and methods for visualizing a pattern in a dataset
GB201704402D0 (en) 2017-03-20 2017-05-03 Blacktrace Holdings Ltd Single cell DNA sequencing
KR20190133711A (ko) 2017-03-24 2019-12-03 싱가포르국립대학교 분자의 다중 검출 방법
US10697008B2 (en) 2017-04-12 2020-06-30 Karius, Inc. Sample preparation methods, systems and compositions
US12054764B2 (en) 2017-04-14 2024-08-06 The Broad Institute, Inc. High-throughput screens for exploring biological functions of microscale biological systems
US20180312822A1 (en) 2017-04-26 2018-11-01 10X Genomics, Inc. Mmlv reverse transcriptase variants
EP4215616B1 (en) 2017-05-18 2024-09-04 10X Genomics, Inc. Methods and systems for sorting droplets and beads
US10544413B2 (en) 2017-05-18 2020-01-28 10X Genomics, Inc. Methods and systems for sorting droplets and beads
WO2018213774A1 (en) 2017-05-19 2018-11-22 10X Genomics, Inc. Systems and methods for analyzing datasets
US10914729B2 (en) 2017-05-22 2021-02-09 The Trustees Of Princeton University Methods for detecting protein binding sequences and tagging nucleic acids
US10400235B2 (en) 2017-05-26 2019-09-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
WO2018226546A1 (en) 2017-06-05 2018-12-13 10X Genomics, Inc. Gaskets for the distribution of pressures in a microfluidic system
AU2018281745B2 (en) 2017-06-05 2022-05-19 Becton, Dickinson And Company Sample indexing for single cells
CN110799679A (zh) 2017-06-20 2020-02-14 10X基因组学有限公司 用于改善液滴稳定的方法和系统
EP3642358A1 (en) 2017-06-21 2020-04-29 Bluedot LLC Systems and methods for identification of nucleic acids in a sample
CN108336542B (zh) 2017-06-23 2020-02-21 番禺得意精密电子工业有限公司 电连接器
NZ759924A (en) 2017-08-01 2023-07-28 Illumina Inc Hydrogel beads for nucleotide sequencing
US9946577B1 (en) 2017-08-14 2018-04-17 10X Genomics, Inc. Systems and methods for distributed resource management
US10821442B2 (en) 2017-08-22 2020-11-03 10X Genomics, Inc. Devices, systems, and kits for forming droplets
EP3691703A1 (en) 2017-10-04 2020-08-12 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US10590244B2 (en) 2017-10-04 2020-03-17 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US10837047B2 (en) 2017-10-04 2020-11-17 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US20190127731A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. Methods for preparing nucleic acid molecules
WO2019083852A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. MICROFLUIDIC CHANNEL NETWORKS FOR PARTITIONING
WO2019084328A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS FOR PREPARING NUCLEIC ACID MOLECULES
WO2019084043A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS AND SYSTEMS FOR NUCLEIC ACID PREPARATION AND CHROMATIN ANALYSIS
EP4241882B1 (en) 2017-10-27 2025-04-30 10X Genomics, Inc. Methods for sample preparation and analysis
CN111051523B (zh) 2017-11-15 2024-03-19 10X基因组学有限公司 功能化凝胶珠
US10829815B2 (en) 2017-11-17 2020-11-10 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
WO2019108851A1 (en) 2017-11-30 2019-06-06 10X Genomics, Inc. Systems and methods for nucleic acid preparation and analysis
WO2019113235A1 (en) 2017-12-06 2019-06-13 10X Genomics, Inc. Methods and systems for processing nucleic acid molecules
CN111699388B (zh) 2017-12-12 2024-08-02 10X基因组学有限公司 用于单细胞处理的系统和方法
CN111712579B (zh) 2017-12-22 2024-10-15 10X基因组学有限公司 用于处理来自一个或多个细胞的核酸分子的系统和方法
CN108173158B (zh) 2018-01-05 2023-09-08 浙江正泰电器股份有限公司 抽屉式断路器
WO2019148042A1 (en) 2018-01-26 2019-08-01 10X Genomics, Inc. Compositions and methods for sample processing
CN112004920B (zh) 2018-02-05 2024-08-02 斯坦福大学托管董事会 用于单细胞和集合细胞的多重测量的系统和方法
SG11202007686VA (en) 2018-02-12 2020-09-29 10X Genomics Inc Methods characterizing multiple analytes from individual cells or cell populations
EP3755812A1 (en) 2018-02-22 2020-12-30 10X Genomics, Inc. Ligation mediated analysis of nucleic acids
WO2019169028A1 (en) 2018-02-28 2019-09-06 10X Genomics, Inc. Transcriptome sequencing through random ligation
WO2019169347A1 (en) 2018-03-02 2019-09-06 10X Genomics, Inc. Systems and apparatus for holding plates
WO2019191321A1 (en) 2018-03-28 2019-10-03 10X Genomics, Inc. Nucleic acid enrichment within partitions
SG11202009889VA (en) 2018-04-06 2020-11-27 10X Genomics Inc Systems and methods for quality control in single cell processing
WO2019217758A1 (en) 2018-05-10 2019-11-14 10X Genomics, Inc. Methods and systems for molecular library generation
US20190345636A1 (en) 2018-05-10 2019-11-14 10X Genomics, Inc. Methods and systems for molecular library generation
US20190352717A1 (en) 2018-05-18 2019-11-21 10X Genomics, Inc. Targeted non-invasive prenatal testing
US11932899B2 (en) 2018-06-07 2024-03-19 10X Genomics, Inc. Methods and systems for characterizing nucleic acid molecules
US11703427B2 (en) 2018-06-25 2023-07-18 10X Genomics, Inc. Methods and systems for cell and bead processing
US11574706B2 (en) 2018-06-28 2023-02-07 10X Genomics, Inc. Systems and methods for visualization of single-cell resolution characteristics
US20200032335A1 (en) 2018-07-27 2020-01-30 10X Genomics, Inc. Systems and methods for metabolome analysis
US20200033366A1 (en) 2018-07-27 2020-01-30 10X Genomics, Inc. Systems and methods for metabolome analysis
SG11202101164TA (en) 2018-08-03 2021-03-30 10X Genomics Inc Methods and systems to minimize barcode exchange
US12065688B2 (en) 2018-08-20 2024-08-20 10X Genomics, Inc. Compositions and methods for cellular processing
WO2020041148A1 (en) 2018-08-20 2020-02-27 10X Genomics, Inc. Methods and systems for detection of protein-dna interactions using proximity ligation
US20200105373A1 (en) 2018-09-28 2020-04-02 10X Genomics, Inc. Systems and methods for cellular analysis using nucleic acid sequencing
US11459607B1 (en) 2018-12-10 2022-10-04 10X Genomics, Inc. Systems and methods for processing-nucleic acid molecules from a single cell using sequential co-partitioning and composite barcodes
EP3906318A1 (en) 2019-01-06 2021-11-10 10X Genomics, Inc. Methods and systems for enrichment of barcodes
WO2020167862A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transfer of reagents between droplets
US11467153B2 (en) 2019-02-12 2022-10-11 10X Genomics, Inc. Methods for processing nucleic acid molecules
WO2020167866A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transposon loading
CN113614245B (zh) 2019-02-12 2024-08-27 10X基因组学有限公司 用于加工核酸分子的方法
US11655499B1 (en) 2019-02-25 2023-05-23 10X Genomics, Inc. Detection of sequence elements in nucleic acid molecules
SG11202111242PA (en) 2019-03-11 2021-11-29 10X Genomics Inc Systems and methods for processing optically tagged beads
EP3947722A1 (en) 2019-03-27 2022-02-09 10X Genomics, Inc. Systems and methods for processing rna from cells
CN114729392A (zh) 2019-09-06 2022-07-08 10X基因组学有限公司 用于对细胞和细胞珠粒进行条形码化的系统和方法
US12241890B2 (en) 2019-12-23 2025-03-04 10X Genomics, Inc. Methods for generating barcoded nucleic acid molecules using fixed cells
WO2021133845A1 (en) 2019-12-23 2021-07-01 10X Genomics, Inc. Reversible fixing reagents and methods of use thereof
CN115461470A (zh) 2020-02-28 2022-12-09 10X基因组学有限公司 用于从组织中分离核和细胞的方法
WO2021207610A1 (en) 2020-04-10 2021-10-14 10X Genomics, Inc. Cold protease treatment method for preparing biological samples
WO2021212042A1 (en) 2020-04-16 2021-10-21 10X Genomics, Inc. Compositions and methods for use with fixed samples
WO2021222302A1 (en) 2020-04-27 2021-11-04 10X Genomics, Inc. Methods and systems for increasing cell recovery efficiency
WO2021222301A1 (en) 2020-04-27 2021-11-04 10X Genomics, Inc. Methods and systems for analysis and identification of barcode multiplets
EP4244379A1 (en) 2020-11-13 2023-09-20 10X Genomics, Inc. Nano-partitions (encapsulated nucleic acid processing enzymes) for cell-lysis and multiple reactions in partition-based assays
EP4298239A1 (en) 2021-02-23 2024-01-03 10X Genomics, Inc. Drug screening methods
WO2022182682A1 (en) 2021-02-23 2022-09-01 10X Genomics, Inc. Probe-based analysis of nucleic acids and proteins
WO2022271908A1 (en) 2021-06-23 2022-12-29 10X Genomics, Inc. Chop-fix method and chopping device for preparing biological samples
EP4419911A4 (en) 2021-10-28 2025-03-05 10X Genomics, Inc. Methods for sample preparation
WO2024006392A1 (en) 2022-06-29 2024-01-04 10X Genomics, Inc. Probe-based analysis of nucleic acids and proteins

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004010106A2 (en) * 2002-07-24 2004-01-29 Ptc Therapeutics, Inc. METHODS FOR IDENTIFYING SMALL MOLEDULES THAT MODULATE PREMATURE TRANSLATION TERMINATION AND NONSENSE MEDIATED mRNA DECAY
WO2006030993A1 (en) * 2004-09-14 2006-03-23 Jin-Ho Choy Information code system using dna sequences
US20090053169A1 (en) * 2007-08-20 2009-02-26 Pharmain Corporation Oligonucleotide Core Carrier Compositions for Delivery of Nucleic Acid-Containing Therapeutic Agents, Methods of Making and Using the Same
WO2010004018A2 (en) * 2008-07-11 2010-01-14 Eth Zurich Degradable microcapsules
WO2011066476A1 (en) * 2009-11-25 2011-06-03 Quantalife, Inc. Methods and compositions for detecting genetic material

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
TUBELEVICIUTE, A ET AL.: "Compartmentalized Self-replication (CSR) Selection Of Thermococcus Litoralis Sh1B DNA Polymerase For Diminished Uracil Binding.", PROTEIN ENGINEERING, DESIGN & SELECTION., vol. 23, no. 8, 31 May 2010 (2010-05-31), pages 589 - 597, XP055186717 *
ZHANG, L ET AL.: "Degradable Disulfide Core-Cross-Linked Micelles As A Drug Delivery System Prepared From Vinyl Functionalized Nucleosides Via The RAFT Process.", BIOMACROMOLECULES., vol. 9, no. 11, 9 October 2008 (2008-10-09), pages 3321 - 3331, XP055187030 *

Cited By (296)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10392661B2 (en) 2009-12-15 2019-08-27 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US11970737B2 (en) 2009-12-15 2024-04-30 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US10619203B2 (en) 2009-12-15 2020-04-14 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US9290809B2 (en) 2009-12-15 2016-03-22 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US9290808B2 (en) 2009-12-15 2016-03-22 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US9315857B2 (en) 2009-12-15 2016-04-19 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse label-tags
US11993814B2 (en) 2009-12-15 2024-05-28 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US9816137B2 (en) 2009-12-15 2017-11-14 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US9708659B2 (en) 2009-12-15 2017-07-18 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US12060607B2 (en) 2009-12-15 2024-08-13 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US10047394B2 (en) 2009-12-15 2018-08-14 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US10202646B2 (en) 2009-12-15 2019-02-12 Becton, Dickinson And Company Digital counting of individual molecules by stochastic attachment of diverse labels
US10059991B2 (en) 2009-12-15 2018-08-28 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US9845502B2 (en) 2009-12-15 2017-12-19 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US11634708B2 (en) 2012-02-27 2023-04-25 Becton, Dickinson And Company Compositions and kits for molecular counting
US11177020B2 (en) 2012-02-27 2021-11-16 The University Of North Carolina At Chapel Hill Methods and uses for molecular tags
US10941396B2 (en) 2012-02-27 2021-03-09 Becton, Dickinson And Company Compositions and kits for molecular counting
US11001883B2 (en) 2012-03-05 2021-05-11 The General Hospital Corporation Systems and methods for epigenetic sequencing
US11047003B2 (en) 2012-03-05 2021-06-29 The General Hospital Corporation Systems and methods for epigenetic sequencing
US11891666B2 (en) 2012-08-13 2024-02-06 The Regents Of The University Of California Methods and systems for detecting biological components
US11001896B2 (en) 2012-08-13 2021-05-11 The Regents Of The University Of California System and method to synthesize a target molecule within a droplet
US10745762B2 (en) 2012-08-13 2020-08-18 The Regents Of The University Of California Method and system for synthesizing a target polynucleotide within a droplet
US11203787B2 (en) 2012-08-13 2021-12-21 The Regents Of The University Of California Methods and systems for detecting biological components
US10450607B2 (en) 2012-08-14 2019-10-22 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9695468B2 (en) 2012-08-14 2017-07-04 10X Genomics, Inc. Methods for droplet-based sample preparation
US11035002B2 (en) 2012-08-14 2021-06-15 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10584381B2 (en) 2012-08-14 2020-03-10 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9689024B2 (en) 2012-08-14 2017-06-27 10X Genomics, Inc. Methods for droplet-based sample preparation
US10323279B2 (en) 2012-08-14 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11021749B2 (en) 2012-08-14 2021-06-01 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10752949B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10273541B2 (en) 2012-08-14 2019-04-30 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10752950B2 (en) 2012-08-14 2020-08-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11078522B2 (en) 2012-08-14 2021-08-03 10X Genomics, Inc. Capsule array devices and methods of use
US10400280B2 (en) 2012-08-14 2019-09-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11441179B2 (en) 2012-08-14 2022-09-13 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10221442B2 (en) 2012-08-14 2019-03-05 10X Genomics, Inc. Compositions and methods for sample processing
US12037634B2 (en) 2012-08-14 2024-07-16 10X Genomics, Inc. Capsule array devices and methods of use
US12098423B2 (en) 2012-08-14 2024-09-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11359239B2 (en) 2012-08-14 2022-06-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10669583B2 (en) 2012-08-14 2020-06-02 10X Genomics, Inc. Method and systems for processing polynucleotides
US10626458B2 (en) 2012-08-14 2020-04-21 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10053723B2 (en) 2012-08-14 2018-08-21 10X Genomics, Inc. Capsule array devices and methods of use
US10597718B2 (en) 2012-08-14 2020-03-24 10X Genomics, Inc. Methods and systems for sample processing polynucleotides
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
US9701998B2 (en) 2012-12-14 2017-07-11 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10227648B2 (en) 2012-12-14 2019-03-12 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11421274B2 (en) 2012-12-14 2022-08-23 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9410201B2 (en) 2012-12-14 2016-08-09 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11473138B2 (en) 2012-12-14 2022-10-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10253364B2 (en) 2012-12-14 2019-04-09 10X Genomics, Inc. Method and systems for processing polynucleotides
US10612090B2 (en) 2012-12-14 2020-04-07 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9856530B2 (en) 2012-12-14 2018-01-02 10X Genomics, Inc. Methods and systems for processing polynucleotides
US9567631B2 (en) 2012-12-14 2017-02-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10676789B2 (en) 2012-12-14 2020-06-09 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10533221B2 (en) 2012-12-14 2020-01-14 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10150963B2 (en) 2013-02-08 2018-12-11 10X Genomics, Inc. Partitioning and processing of analytes and other species
US11193121B2 (en) 2013-02-08 2021-12-07 10X Genomics, Inc. Partitioning and processing of analytes and other species
US10150964B2 (en) 2013-02-08 2018-12-11 10X Genomics, Inc. Partitioning and processing of analytes and other species
US9388465B2 (en) 2013-02-08 2016-07-12 10X Genomics, Inc. Polynucleotide barcode generation
US9644204B2 (en) 2013-02-08 2017-05-09 10X Genomics, Inc. Partitioning and processing of analytes and other species
EP3013957B1 (en) 2013-06-27 2018-09-26 10X Genomics, Inc. Compositions and methods for sample processing
US9567645B2 (en) 2013-08-28 2017-02-14 Cellular Research, Inc. Massively parallel single cell analysis
US9637799B2 (en) 2013-08-28 2017-05-02 Cellular Research, Inc. Massively parallel single cell analysis
US10927419B2 (en) 2013-08-28 2021-02-23 Becton, Dickinson And Company Massively parallel single cell analysis
US10954570B2 (en) 2013-08-28 2021-03-23 Becton, Dickinson And Company Massively parallel single cell analysis
US10151003B2 (en) 2013-08-28 2018-12-11 Cellular Research, Inc. Massively Parallel single cell analysis
US10131958B1 (en) 2013-08-28 2018-11-20 Cellular Research, Inc. Massively parallel single cell analysis
US9598736B2 (en) 2013-08-28 2017-03-21 Cellular Research, Inc. Massively parallel single cell analysis
US10208356B1 (en) 2013-08-28 2019-02-19 Becton, Dickinson And Company Massively parallel single cell analysis
US10253375B1 (en) 2013-08-28 2019-04-09 Becton, Dickinson And Company Massively parallel single cell analysis
US11702706B2 (en) 2013-08-28 2023-07-18 Becton, Dickinson And Company Massively parallel single cell analysis
US11618929B2 (en) 2013-08-28 2023-04-04 Becton, Dickinson And Company Massively parallel single cell analysis
US9567646B2 (en) 2013-08-28 2017-02-14 Cellular Research, Inc. Massively parallel single cell analysis
US10395758B2 (en) 2013-08-30 2019-08-27 10X Genomics, Inc. Sequencing methods
US12249402B2 (en) 2013-08-30 2025-03-11 10X Genomics, Inc. Sequencing methods
US12131805B2 (en) 2013-08-30 2024-10-29 10X Genomics, Inc. Sequencing methods
US12006543B2 (en) 2013-09-13 2024-06-11 Life Technologies Corporation Device preparation using condensed nucleic acid particles
US9905005B2 (en) 2013-10-07 2018-02-27 Cellular Research, Inc. Methods and systems for digitally counting features on arrays
US9582877B2 (en) 2013-10-07 2017-02-28 Cellular Research, Inc. Methods and systems for digitally counting features on arrays
US10947587B2 (en) * 2013-11-05 2021-03-16 The Regents Of The University Of California Single-cell forensic short tandem repeat typing within microfluidic droplets
US11030276B2 (en) 2013-12-16 2021-06-08 10X Genomics, Inc. Methods and apparatus for sorting data
US11853389B2 (en) 2013-12-16 2023-12-26 10X Genomics, Inc. Methods and apparatus for sorting data
US9824068B2 (en) 2013-12-16 2017-11-21 10X Genomics, Inc. Methods and apparatus for sorting data
US9694361B2 (en) 2014-04-10 2017-07-04 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
EP4219010A1 (en) 2014-04-10 2023-08-02 10X Genomics, Inc. Methods for encapsulating and partitioning reagents
CN106413896B (zh) * 2014-04-10 2019-07-05 10X基因组学有限公司 用于封装和分割试剂的流体装置、系统和方法及其应用
US12005454B2 (en) 2014-04-10 2024-06-11 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
CN114534806B (zh) * 2014-04-10 2024-03-29 10X基因组学有限公司 用于封装和分割试剂的流体装置、系统和方法及其应用
CN106413896A (zh) * 2014-04-10 2017-02-15 10X基因组学有限公司 用于封装和分割试剂的流体装置、系统和方法及其应用
WO2015157567A1 (en) 2014-04-10 2015-10-15 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
US10071377B2 (en) 2014-04-10 2018-09-11 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
KR102596508B1 (ko) 2014-04-10 2023-10-30 10엑스 제노믹스, 인크. 시약을 캡슐화 및 구획화하기 위한 유체 디바이스, 시스템, 및 방법, 및 이의 응용
US10137449B2 (en) 2014-04-10 2018-11-27 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
KR20160142883A (ko) 2014-04-10 2016-12-13 10엑스 제노믹스, 인크. 시약을 캡슐화 및 구획화하기 위한 유체 디바이스, 시스템, 및 방법, 및 이의 응용
CN114534806A (zh) * 2014-04-10 2022-05-27 10X基因组学有限公司 用于封装和分割试剂的流体装置、系统和方法及其应用
AU2015243445B2 (en) * 2014-04-10 2020-05-28 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
DE202015009494U1 (de) 2014-04-10 2018-02-08 10X Genomics, Inc. Fluidische Vorrichtungen und Systeme zur Einkapselung und Partitionierung von Reagenzien, und deren Anwendungen
US10150117B2 (en) 2014-04-10 2018-12-11 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
CN110548550A (zh) * 2014-04-10 2019-12-10 10X基因组学有限公司 用于封装和分割试剂的流体装置、系统和方法及其应用
US10343166B2 (en) 2014-04-10 2019-07-09 10X Genomics, Inc. Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same
US10518241B2 (en) * 2014-04-15 2019-12-31 Agilent Technologies, Inc. Creating and harvesting surface-bound emulsion
US10596541B2 (en) 2014-04-21 2020-03-24 President And Fellows Of Harvard College Systems and methods for barcoding nucleic acids
US11052368B2 (en) 2014-04-21 2021-07-06 Vilnius University Systems and methods for barcoding nucleic acids
US11155809B2 (en) 2014-06-24 2021-10-26 Bio-Rad Laboratories, Inc. Digital PCR barcoding
US11629344B2 (en) 2014-06-26 2023-04-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
CN106795553B (zh) * 2014-06-26 2021-06-04 10X基因组学有限公司 分析来自单个细胞或细胞群体的核酸的方法
US10208343B2 (en) 2014-06-26 2019-02-19 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10041116B2 (en) 2014-06-26 2018-08-07 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10030267B2 (en) 2014-06-26 2018-07-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
US12312640B2 (en) 2014-06-26 2025-05-27 10X Genomics, Inc. Analysis of nucleic acid sequences
US10480028B2 (en) 2014-06-26 2019-11-19 10X Genomics, Inc. Methods and systems for processing polynucleotides
WO2015200871A1 (en) * 2014-06-26 2015-12-30 10X Genomics, Inc. Methods and compositions for sample analysis
US11133084B2 (en) 2014-06-26 2021-09-28 10X Genomics, Inc. Systems and methods for nucleic acid sequence assembly
US12163191B2 (en) 2014-06-26 2024-12-10 10X Genomics, Inc. Analysis of nucleic acid sequences
US10457986B2 (en) 2014-06-26 2019-10-29 10X Genomics, Inc. Methods and systems for processing polynucleotides
CN113249435A (zh) * 2014-06-26 2021-08-13 10X基因组学有限公司 分析来自单个细胞或细胞群体的核酸的方法
US20150376605A1 (en) * 2014-06-26 2015-12-31 10X Genomics, Inc. Methods and Compositions for Sample Analysis
US9951386B2 (en) 2014-06-26 2018-04-24 10X Genomics, Inc. Methods and systems for processing polynucleotides
KR102531677B1 (ko) 2014-06-26 2023-05-10 10엑스 제노믹스, 인크. 개별 세포 또는 세포 개체군으로부터 핵산을 분석하는 방법
JP2017522867A (ja) * 2014-06-26 2017-08-17 １０エックス ジェノミクス， インコーポレイテッド 個々の細胞または細胞集団由来の核酸の分析方法
US10839939B2 (en) 2014-06-26 2020-11-17 10X Genomics, Inc. Processes and systems for nucleic acid sequence assembly
JP2017522866A (ja) * 2014-06-26 2017-08-17 １０エックス ジェノミクス， インコーポレイテッド 核酸配列の分析
US10337061B2 (en) 2014-06-26 2019-07-02 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11713457B2 (en) 2014-06-26 2023-08-01 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10344329B2 (en) 2014-06-26 2019-07-09 10X Genomics, Inc. Methods and systems for processing polynucleotides
KR20170020704A (ko) * 2014-06-26 2017-02-23 10엑스 제노믹스, 인크. 개별 세포 또는 세포 개체군으로부터 핵산을 분석하는 방법
CN106574298A (zh) * 2014-06-26 2017-04-19 10X基因组学有限公司 用于样品分析的方法和组合物
US10760124B2 (en) 2014-06-26 2020-09-01 10X Genomics, Inc. Methods and systems for processing polynucleotides
EP3161162A4 (en) * 2014-06-26 2018-01-10 10X Genomics, Inc. Analysis of nucleic acid sequences
CN106795553A (zh) * 2014-06-26 2017-05-31 10X基因组学有限公司 分析来自单个细胞或细胞群体的核酸的方法
US10697007B2 (en) 2014-06-27 2020-06-30 The Regents Of The University Of California PCR-activated sorting (PAS)
US11312990B2 (en) 2014-06-27 2022-04-26 The Regents Of The University Of California PCR-activated sorting (PAS)
JP2017522049A (ja) * 2014-07-02 2017-08-10 ライフ テクノロジーズ コーポレーション センサー基質を装填するための方法
US11020736B2 (en) 2014-10-22 2021-06-01 The Regents Of The University Of California High definition microdroplet printer
US10434507B2 (en) 2014-10-22 2019-10-08 The Regents Of The University Of California High definition microdroplet printer
US10287623B2 (en) 2014-10-29 2019-05-14 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequencing
US11739368B2 (en) 2014-10-29 2023-08-29 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequencing
US11135584B2 (en) 2014-11-05 2021-10-05 10X Genomics, Inc. Instrument systems for integrated sample processing
US10245587B2 (en) 2014-11-05 2019-04-02 10X Genomics, Inc. Instrument systems for integrated sample processing
US9975122B2 (en) 2014-11-05 2018-05-22 10X Genomics, Inc. Instrument systems for integrated sample processing
US10221436B2 (en) 2015-01-12 2019-03-05 10X Genomics, Inc. Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same
US11414688B2 (en) 2015-01-12 2022-08-16 10X Genomics, Inc. Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same
US10557158B2 (en) 2015-01-12 2020-02-11 10X Genomics, Inc. Processes and systems for preparation of nucleic acid sequencing libraries and libraries prepared using same
US12387821B2 (en) 2015-01-13 2025-08-12 10X Genomics, Inc. Systems and methods for visualizing structural variation and phasing information
US10650912B2 (en) 2015-01-13 2020-05-12 10X Genomics, Inc. Systems and methods for visualizing structural variation and phasing information
US11111519B2 (en) 2015-02-04 2021-09-07 The Regents Of The University Of California Sequencing of nucleic acids via barcoding in discrete entities
US11732287B2 (en) 2015-02-04 2023-08-22 The Regents Of The University Of California Sequencing of nucleic acids via barcoding in discrete entities
US10854315B2 (en) 2015-02-09 2020-12-01 10X Genomics, Inc. Systems and methods for determining structural variation and phasing using variant call data
US11098358B2 (en) 2015-02-19 2021-08-24 Becton, Dickinson And Company High-throughput single-cell analysis combining proteomic and genomic information
US10697010B2 (en) 2015-02-19 2020-06-30 Becton, Dickinson And Company High-throughput single-cell analysis combining proteomic and genomic information
US10697000B2 (en) 2015-02-24 2020-06-30 10X Genomics, Inc. Partition processing methods and systems
US20160244809A1 (en) * 2015-02-24 2016-08-25 10X Genomics, Inc. Partition Processing Methods and Systems
US11603554B2 (en) 2015-02-24 2023-03-14 10X Genomics, Inc. Partition processing methods and systems
US11274343B2 (en) 2015-02-24 2022-03-15 10X Genomics, Inc. Methods and compositions for targeted nucleic acid sequence coverage
US10002316B2 (en) 2015-02-27 2018-06-19 Cellular Research, Inc. Spatially addressable molecular barcoding
USRE48913E1 (en) 2015-02-27 2022-02-01 Becton, Dickinson And Company Spatially addressable molecular barcoding
US9727810B2 (en) 2015-02-27 2017-08-08 Cellular Research, Inc. Spatially addressable molecular barcoding
EP3929291A1 (en) 2015-03-17 2021-12-29 Bio-Rad Laboratories, Inc. Detection of genome editing
WO2016149547A1 (en) 2015-03-17 2016-09-22 Bio-Rad Laboratories, Inc. Detection of genome editing
EP3271713A4 (en) * 2015-03-18 2018-10-24 The Broad Institute Inc. Massively parallel on-chip coalescence of microemulsions
US10981167B2 (en) 2015-03-18 2021-04-20 The Broad Institute, Inc. Massively parallel on-chip coalescence of microemulsions
US11535882B2 (en) 2015-03-30 2022-12-27 Becton, Dickinson And Company Methods and compositions for combinatorial barcoding
US11746367B2 (en) 2015-04-17 2023-09-05 President And Fellows Of Harvard College Barcoding systems and methods for gene sequencing and other applications
US11104939B2 (en) 2015-04-21 2021-08-31 General Automation Lab Technologies Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
CN107810277B (zh) * 2015-04-21 2021-11-02 通用自动化实验技术公司 用于高通量微生物学应用的高分辨率系统、试剂盒、设备和方法
US10900073B2 (en) 2015-04-21 2021-01-26 General Automation Lab Technologies Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
WO2016172362A1 (en) * 2015-04-21 2016-10-27 General Automation Lab Technologies, Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
IL287247B1 (en) * 2015-04-21 2023-03-01 Isolation Bio Inc High-resolution systems, kits, devices and methods for high-throughput microbiology applications
CN107810277A (zh) * 2015-04-21 2018-03-16 通用自动化实验技术公司 用于高通量微生物学应用的高分辨率系统、试剂盒、设备和方法
IL287247B2 (en) * 2015-04-21 2023-07-01 Isolation Bio Inc High-resolution systems, kits, devices and methods for high-throughput microbiology applications
US11136621B2 (en) 2015-04-21 2021-10-05 General Automation Lab Technologies Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
US12123888B2 (en) 2015-04-21 2024-10-22 Isolation Bio Inc. Apparatus and method for picking biological sample
IL254858B (en) * 2015-04-21 2021-12-01 Gen Automation Lab Tech Inc High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
EP4070887A1 (en) * 2015-04-21 2022-10-12 Isolation Bio Inc. Methods for high throughput microbiology applications
US10227640B2 (en) 2015-04-21 2019-03-12 General Automation Lab Technologies, Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
US11104940B2 (en) 2015-04-21 2021-08-31 General Automation Lab Technologies Inc. High resolution systems, kits, apparatus, and methods for high throughput microbiology applications
US11390914B2 (en) 2015-04-23 2022-07-19 Becton, Dickinson And Company Methods and compositions for whole transcriptome amplification
US11124823B2 (en) 2015-06-01 2021-09-21 Becton, Dickinson And Company Methods for RNA quantification
US10619186B2 (en) 2015-09-11 2020-04-14 Cellular Research, Inc. Methods and compositions for library normalization
US11332776B2 (en) 2015-09-11 2022-05-17 Becton, Dickinson And Company Methods and compositions for library normalization
US11331417B2 (en) 2015-09-29 2022-05-17 Shanghai Clinical Engine Technology Development Co., Ltd. Magnetic target separation instrument and application thereof
US10774370B2 (en) 2015-12-04 2020-09-15 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
US11624085B2 (en) 2015-12-04 2023-04-11 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
US11473125B2 (en) 2015-12-04 2022-10-18 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
US11873528B2 (en) 2015-12-04 2024-01-16 10X Genomics, Inc. Methods and compositions for nucleic acid analysis
US11081208B2 (en) 2016-02-11 2021-08-03 10X Genomics, Inc. Systems, methods, and media for de novo assembly of whole genome sequence data
US10233487B2 (en) * 2016-03-24 2019-03-19 Bio-Rad Laboratories, Inc. Use of gel beads to control droplet dispersion
US10822643B2 (en) 2016-05-02 2020-11-03 Cellular Research, Inc. Accurate molecular barcoding
US12138628B2 (en) 2016-05-13 2024-11-12 10X Genomics, Inc. Microfluidic systems and methods of use
US11084036B2 (en) 2016-05-13 2021-08-10 10X Genomics, Inc. Microfluidic systems and methods of use
US11845986B2 (en) 2016-05-25 2023-12-19 Becton, Dickinson And Company Normalization of nucleic acid libraries
US10301677B2 (en) 2016-05-25 2019-05-28 Cellular Research, Inc. Normalization of nucleic acid libraries
US11397882B2 (en) 2016-05-26 2022-07-26 Becton, Dickinson And Company Molecular label counting adjustment methods
US11220685B2 (en) 2016-05-31 2022-01-11 Becton, Dickinson And Company Molecular indexing of internal sequences
US10202641B2 (en) 2016-05-31 2019-02-12 Cellular Research, Inc. Error correction in amplification of samples
US11525157B2 (en) 2016-05-31 2022-12-13 Becton, Dickinson And Company Error correction in amplification of samples
US12331351B2 (en) 2016-05-31 2025-06-17 Becton, Dickinson And Company Error correction in amplification of samples
US10640763B2 (en) 2016-05-31 2020-05-05 Cellular Research, Inc. Molecular indexing of internal sequences
EP4435116A2 (en) 2016-08-05 2024-09-25 Bio-Rad Laboratories, Inc. Second strand direct
WO2018027048A1 (en) 2016-08-05 2018-02-08 Bio-Rad Laboratories, Inc. Second strand direct
US11142791B2 (en) 2016-08-10 2021-10-12 The Regents Of The University Of California Combined multiple-displacement amplification and PCR in an emulsion microdroplet
US10688494B2 (en) 2016-08-23 2020-06-23 10X Genomics, Inc. Microfluidic surface-mediated emulsion stability control
US11278901B2 (en) 2016-08-23 2022-03-22 10X Genomics, Inc. Microfluidic surface-mediated emulsion stability control
US11467157B2 (en) 2016-09-26 2022-10-11 Becton, Dickinson And Company Measurement of protein expression using reagents with barcoded oligonucleotide sequences
US10338066B2 (en) 2016-09-26 2019-07-02 Cellular Research, Inc. Measurement of protein expression using reagents with barcoded oligonucleotide sequences
US11460468B2 (en) 2016-09-26 2022-10-04 Becton, Dickinson And Company Measurement of protein expression using reagents with barcoded oligonucleotide sequences
US11782059B2 (en) 2016-09-26 2023-10-10 Becton, Dickinson And Company Measurement of protein expression using reagents with barcoded oligonucleotide sequences
US11608497B2 (en) 2016-11-08 2023-03-21 Becton, Dickinson And Company Methods for cell label classification
US11164659B2 (en) 2016-11-08 2021-11-02 Becton, Dickinson And Company Methods for expression profile classification
WO2018118971A1 (en) 2016-12-19 2018-06-28 Bio-Rad Laboratories, Inc. Droplet tagging contiguity preserved tagmented dna
US11124830B2 (en) 2016-12-21 2021-09-21 The Regents Of The University Of California Single cell genomic sequencing using hydrogel based droplets
US10793905B2 (en) 2016-12-22 2020-10-06 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11180805B2 (en) 2016-12-22 2021-11-23 10X Genomics, Inc Methods and systems for processing polynucleotides
US10480029B2 (en) 2016-12-22 2019-11-19 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10011872B1 (en) 2016-12-22 2018-07-03 10X Genomics, Inc. Methods and systems for processing polynucleotides
US12084716B2 (en) 2016-12-22 2024-09-10 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10323278B2 (en) 2016-12-22 2019-06-18 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10550429B2 (en) 2016-12-22 2020-02-04 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10858702B2 (en) 2016-12-22 2020-12-08 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10815525B2 (en) 2016-12-22 2020-10-27 10X Genomics, Inc. Methods and systems for processing polynucleotides
US10722880B2 (en) 2017-01-13 2020-07-28 Cellular Research, Inc. Hydrophilic coating of fluidic channels
US12264316B2 (en) 2017-01-30 2025-04-01 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
US10428326B2 (en) 2017-01-30 2019-10-01 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
US11193122B2 (en) 2017-01-30 2021-12-07 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
US12264411B2 (en) 2017-01-30 2025-04-01 10X Genomics, Inc. Methods and systems for analysis
US11319583B2 (en) 2017-02-01 2022-05-03 Becton, Dickinson And Company Selective amplification using blocking oligonucleotides
US11898206B2 (en) 2017-05-19 2024-02-13 10X Genomics, Inc. Systems and methods for clonotype screening
US11773389B2 (en) 2017-05-26 2023-10-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US10927370B2 (en) 2017-05-26 2021-02-23 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US10400235B2 (en) 2017-05-26 2019-09-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US11198866B2 (en) 2017-05-26 2021-12-14 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US11155810B2 (en) 2017-05-26 2021-10-26 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US10844372B2 (en) 2017-05-26 2020-11-24 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
US10669570B2 (en) 2017-06-05 2020-06-02 Becton, Dickinson And Company Sample indexing for single cells
US12084712B2 (en) 2017-06-05 2024-09-10 Becton, Dickinson And Company Sample indexing for single cells
US12371729B2 (en) 2017-06-05 2025-07-29 Becton, Dickinson And Company Sample indexing for single cells
US10676779B2 (en) 2017-06-05 2020-06-09 Becton, Dickinson And Company Sample indexing for single cells
US11781129B2 (en) 2017-10-18 2023-10-10 Mission Bio, Inc. Method, systems and apparatus for single cell analysis
US10501739B2 (en) 2017-10-18 2019-12-10 Mission Bio, Inc. Method, systems and apparatus for single cell analysis
EP4180534A1 (en) 2017-11-02 2023-05-17 Bio-Rad Laboratories, Inc. Transposase-based genomic analysis
WO2019089959A1 (en) 2017-11-02 2019-05-09 Bio-Rad Laboratories, Inc. Transposase-based genomic analysis
US10745742B2 (en) 2017-11-15 2020-08-18 10X Genomics, Inc. Functionalized gel beads
US10876147B2 (en) 2017-11-15 2020-12-29 10X Genomics, Inc. Functionalized gel beads
US11884962B2 (en) 2017-11-15 2024-01-30 10X Genomics, Inc. Functionalized gel beads
US10829815B2 (en) 2017-11-17 2020-11-10 10X Genomics, Inc. Methods and systems for associating physical and genetic properties of biological particles
US11946095B2 (en) 2017-12-19 2024-04-02 Becton, Dickinson And Company Particles associated with oligonucleotides
CN111712579A (zh) * 2017-12-22 2020-09-25 10X基因组学有限公司 用于处理来自一个或多个细胞的核酸分子的系统和方法
WO2019126789A1 (en) * 2017-12-22 2019-06-27 10X Genomics, Inc. Systems and methods for processing nucleic acid molecules from one or more cells
US12104200B2 (en) 2017-12-22 2024-10-01 10X Genomics, Inc Systems and methods for processing nucleic acid molecules from one or more cells
EP3505933A1 (en) * 2017-12-29 2019-07-03 Blink AG A microcapsule for detecting and/or quantitating an analyte in a sample
WO2019129580A1 (en) * 2017-12-29 2019-07-04 Blink Ag A microcapsule for detecting and/or quantitating an analyte in a sample
WO2019148042A1 (en) * 2018-01-26 2019-08-01 10X Genomics, Inc. Compositions and methods for sample processing
EP4324962A2 (en) 2018-01-31 2024-02-21 Bio-Rad Laboratories, Inc. Methods and compositions for deconvoluting partition barcodes
WO2019152395A1 (en) 2018-01-31 2019-08-08 Bio-Rad Laboratories, Inc. Methods and compositions for deconvoluting partition barcodes
US11155881B2 (en) 2018-04-06 2021-10-26 10X Genomics, Inc. Systems and methods for quality control in single cell processing
US11773441B2 (en) 2018-05-03 2023-10-03 Becton, Dickinson And Company High throughput multiomics sample analysis
US11365409B2 (en) 2018-05-03 2022-06-21 Becton, Dickinson And Company Molecular barcoding on opposite transcript ends
EP3797279A4 (en) * 2018-07-11 2022-08-03 The University of Hong Kong Automatic microfluidic system for continuous and quantitive collection of droplets
EP4249651A2 (en) 2018-08-20 2023-09-27 Bio-Rad Laboratories, Inc. Nucleotide sequence generation by barcode bead-colocalization in partitions
WO2020041293A1 (en) 2018-08-20 2020-02-27 Bio-Rad Laboratories, Inc. Nucleotide sequence generation by barcode bead-colocalization in partitions
US11639517B2 (en) 2018-10-01 2023-05-02 Becton, Dickinson And Company Determining 5′ transcript sequences
US11932849B2 (en) 2018-11-08 2024-03-19 Becton, Dickinson And Company Whole transcriptome analysis of single cells using random priming
US11492660B2 (en) 2018-12-13 2022-11-08 Becton, Dickinson And Company Selective extension in single cell whole transcriptome analysis
US11371076B2 (en) 2019-01-16 2022-06-28 Becton, Dickinson And Company Polymerase chain reaction normalization through primer titration
US11661631B2 (en) 2019-01-23 2023-05-30 Becton, Dickinson And Company Oligonucleotides associated with antibodies
US12071617B2 (en) 2019-02-14 2024-08-27 Becton, Dickinson And Company Hybrid targeted and whole transcriptome amplification
US11965208B2 (en) 2019-04-19 2024-04-23 Becton, Dickinson And Company Methods of associating phenotypical data and single cell sequencing data
US11365441B2 (en) 2019-05-22 2022-06-21 Mission Bio, Inc. Method and apparatus for simultaneous targeted sequencing of DNA, RNA and protein
US11667954B2 (en) 2019-07-01 2023-06-06 Mission Bio, Inc. Method and apparatus to normalize quantitative readouts in single-cell experiments
US11939622B2 (en) 2019-07-22 2024-03-26 Becton, Dickinson And Company Single cell chromatin immunoprecipitation sequencing assay
US11773436B2 (en) 2019-11-08 2023-10-03 Becton, Dickinson And Company Using random priming to obtain full-length V(D)J information for immune repertoire sequencing
US11773452B2 (en) 2020-01-13 2023-10-03 Fluent Biosciences Inc. Single cell sequencing
US11512337B2 (en) 2020-01-13 2022-11-29 Fluent Biosciences Inc. Emulsion based drug screening
WO2021146184A1 (en) * 2020-01-13 2021-07-22 Fluent Biosciences Inc. Single cell sequencing
US11649497B2 (en) 2020-01-13 2023-05-16 Becton, Dickinson And Company Methods and compositions for quantitation of proteins and RNA
US11104961B2 (en) 2020-01-13 2021-08-31 Fluent Biosciences Inc. Single cell sequencing
US11827936B2 (en) 2020-01-13 2023-11-28 Fluent Biosciences Inc. Methods and systems for single cell gene profiling
US12188010B2 (en) 2020-01-29 2025-01-07 Becton, Dickinson And Company Barcoded wells for spatial mapping of single cells through sequencing
US11738341B2 (en) 2020-02-10 2023-08-29 Roche Molecular Systems, Inc. Dispensing of highly viscous liquids
EP3862090A1 (en) * 2020-02-10 2021-08-11 Roche Diagnostics GmbH Liquid dispensing system for a microfluidic sample carrier, microfluidic sample carrier sealing system including such liquid dispensing system, and method for dispensing sealing liquid using the same
US12153043B2 (en) 2020-02-25 2024-11-26 Becton, Dickinson And Company Bi-specific probes to enable the use of single-cell samples as single color compensation control
US11866782B2 (en) 2020-03-16 2024-01-09 Fluent Biosciences Inc. Multi-omic analysis in monodisperse droplets
US12378594B2 (en) 2020-05-14 2025-08-05 Becton, Dickinson And Company Primers for immune repertoire profiling
US11661625B2 (en) 2020-05-14 2023-05-30 Becton, Dickinson And Company Primers for immune repertoire profiling
US12157913B2 (en) 2020-06-02 2024-12-03 Becton, Dickinson And Company Oligonucleotides and beads for 5 prime gene expression assay
US11932901B2 (en) 2020-07-13 2024-03-19 Becton, Dickinson And Company Target enrichment using nucleic acid probes for scRNAseq
US12241059B2 (en) 2020-07-15 2025-03-04 Illumina, Inc. Tiered ligation oligos
US12391940B2 (en) 2020-07-31 2025-08-19 Becton, Dickinson And Company Single cell assay for transposase-accessible chromatin
US11739443B2 (en) 2020-11-20 2023-08-29 Becton, Dickinson And Company Profiling of highly expressed and lowly expressed proteins
US12392771B2 (en) 2020-12-15 2025-08-19 Becton, Dickinson And Company Single cell secretome analysis
US12421558B2 (en) 2021-02-12 2025-09-23 10X Genomics, Inc. Systems and methods for joint interactive visualization of gene expression and DNA chromatin accessibility
CN113736850A (zh) * 2021-08-13 2021-12-03 纳昂达(南京)生物科技有限公司 基于双链环化的文库构建方法及其在测序中的应用
US12421548B2 (en) 2023-05-26 2025-09-23 Becton, Dickinson And Company High throughput multiomics sample analysis
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US12421539B2 (en) 2023-11-27 2025-09-23 10X Genomics, Inc. Methods and compositions for nucleic acid analysis

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