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WO2012156744A3 - Gel beads in microfluidic droplets - Google Patents

Gel beads in microfluidic droplets Download PDF

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Publication number
WO2012156744A3
WO2012156744A3 PCT/GB2012/051106 GB2012051106W WO2012156744A3 WO 2012156744 A3 WO2012156744 A3 WO 2012156744A3 GB 2012051106 W GB2012051106 W GB 2012051106W WO 2012156744 A3 WO2012156744 A3 WO 2012156744A3
Authority
WO
WIPO (PCT)
Prior art keywords
reporter
microdroplets
gel
aqueous
beads
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2012/051106
Other languages
French (fr)
Other versions
WO2012156744A2 (en
Inventor
Florian Hollfelder
Martin Fischlechner
Yolanda SCHAERLI
Christopher Abell
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cambridge Enterprise Ltd
Original Assignee
Cambridge Enterprise Ltd
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Filing date
Publication date
Application filed by Cambridge Enterprise Ltd filed Critical Cambridge Enterprise Ltd
Publication of WO2012156744A2 publication Critical patent/WO2012156744A2/en
Publication of WO2012156744A3 publication Critical patent/WO2012156744A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12N15/1075—Isolating an individual clone by screening libraries by coupling phenotype to genotype, not provided for in other groups of this subclass
    • 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
    • C12N15/1086—Preparation or screening of expression libraries, e.g. reporter assays

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

This invention relates to the use of gel beads in microfluidic droplets to perform multi-step compartmentalised reactions in vitro. Methods may comprise emulsifying an aqueous reporter solution which comprises polynucleotides, a reporter substrate, and a gel-forming agent into microdroplets. Each polynucleotide encodes a product which converts the reporter substrate into a detectable reporter in the microdroplets. The gel-forming agent is then solidified within the microdroplets to produce gel beads comprising both the polynucleotide and the detectable reporter produced by the product. The aqueous microdroplets are then de-emulsified and re-suspended in aqueous detection solution and the reporter detected in one or more beads the population. These methods allow high throughput and are particularly useful for biochemical processes such as high-throughput screening, directed evolution and genotyping methods.
PCT/GB2012/051106 2011-05-17 2012-05-17 Gel beads in microfluidic droplets Ceased WO2012156744A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1108259.1A GB201108259D0 (en) 2011-05-17 2011-05-17 Gel beads in microfluidic droplets
GB1108259.1 2011-05-17

Publications (2)

Publication Number Publication Date
WO2012156744A2 WO2012156744A2 (en) 2012-11-22
WO2012156744A3 true WO2012156744A3 (en) 2013-01-10

Family

ID=44260665

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2012/051106 Ceased WO2012156744A2 (en) 2011-05-17 2012-05-17 Gel beads in microfluidic droplets

Country Status (2)

Country Link
GB (1) GB201108259D0 (en)
WO (1) WO2012156744A2 (en)

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US11667954B2 (en) 2019-07-01 2023-06-06 Mission Bio, Inc. Method and apparatus to normalize quantitative readouts in single-cell experiments
US11845986B2 (en) 2016-05-25 2023-12-19 Becton, Dickinson And Company Normalization of nucleic acid libraries

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WO2016172373A1 (en) 2015-04-23 2016-10-27 Cellular Research, Inc. Methods and compositions for whole transcriptome amplification
ES2745694T3 (en) 2015-09-11 2020-03-03 Cellular Res Inc Methods and compositions for nucleic acid library normalization
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CN108883031B (en) * 2016-03-24 2022-01-25 生物辐射实验室股份有限公司 Use of gel beads to control droplet dispersion
WO2017184707A1 (en) * 2016-04-19 2017-10-26 President And Fellows Of Harvard College Immobilization-based systems and methods for genetic analysis and other applications
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US10640763B2 (en) 2016-05-31 2020-05-05 Cellular Research, Inc. Molecular indexing of internal sequences
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WO2018191701A1 (en) * 2017-04-14 2018-10-18 The Broad Institute, Inc. High-throughput screens for exploring biological functions of microscale biological systems
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US10501739B2 (en) * 2017-10-18 2019-12-10 Mission Bio, Inc. Method, systems and apparatus for single cell analysis
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JP7407128B2 (en) 2018-05-03 2023-12-28 ベクトン・ディキンソン・アンド・カンパニー High-throughput multi-omics sample analysis
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US20210382061A1 (en) * 2018-10-22 2021-12-09 The General Hospital Corporation Multiplexed single-cell analysis using optically-encoded rna capture particles
CN112969789A (en) 2018-11-08 2021-06-15 贝克顿迪金森公司 Single cell whole transcriptome analysis using random priming
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CN113574178B (en) 2019-01-23 2024-10-29 贝克顿迪金森公司 Oligonucleotides linked to antibodies
EP3924506A1 (en) 2019-02-14 2021-12-22 Becton Dickinson and Company Hybrid targeted and whole transcriptome amplification
EP3930900A1 (en) 2019-02-28 2022-01-05 10X Genomics, Inc. Devices, systems, and methods for increasing droplet formation efficiency
US11965208B2 (en) 2019-04-19 2024-04-23 Becton, Dickinson And Company Methods of associating phenotypical data and single cell sequencing data
FR3098128B1 (en) * 2019-07-05 2023-11-17 Commissariat Energie Atomique Microfluidic device comprising a microdrop having a sol-gel matrix.
WO2021016239A1 (en) 2019-07-22 2021-01-28 Becton, Dickinson And Company Single cell chromatin immunoprecipitation sequencing assay
ES2987640T3 (en) 2019-11-08 2024-11-15 Becton Dickinson Co Using random priming to obtain full-length V(D)J information for immune repertoire sequencing
WO2021097301A1 (en) 2019-11-15 2021-05-20 Christopher Gordon Atwood Compositions and methods based on diffusion of fluorophores
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11845986B2 (en) 2016-05-25 2023-12-19 Becton, Dickinson And Company Normalization of nucleic acid libraries
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

Also Published As

Publication number Publication date
WO2012156744A2 (en) 2012-11-22
GB201108259D0 (en) 2011-06-29

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