DK2652155T3 - Fremgangsmåder til massiv parallelanalyse af nukleinsyrer i enkeltceller - Google Patents

Fremgangsmåder til massiv parallelanalyse af nukleinsyrer i enkeltceller Download PDF

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DK2652155T3
DK2652155T3 DK11848932.7T DK11848932T DK2652155T3 DK 2652155 T3 DK2652155 T3 DK 2652155T3 DK 11848932 T DK11848932 T DK 11848932T DK 2652155 T3 DK2652155 T3 DK 2652155T3
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sequence
nucleic acid
target nucleic
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probe
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David Scott Johnson
Everett Hurteau Meyer
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Gigagen Inc
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Claims (23)

1. Fremgangsmåde til analyse af mindst to nukleinsyresekvenser i en enkeltcelle, der er indeholdt i en population af mindst 10.000 celler, hvor enkeltcellen isoleres i en emulsionsmikrodråbe, hvilken fremgangsmåde omfatter: tilvejebringelse af et første sæt af nukleinsyreprober, hvilket første sæt omfatter en første probe, der omfatter en sekvens, der er komplementær til en første målnukleinsyreundersekvens, en anden probe, der omfatter en sekvens, der er komplementærtil en anden undersekvens af den første målnukleinsyre, og en anden sekvens, der er komplementær til en eksogen sekvens, en tredje probe, der omfatter den eksogene sekvens og en sekvens, der er komplementær til en første undersekvens af en anden målnukleinsyre, og en Ijerde probe, der omfatter en sekvens, der er komplementær til en anden undersekvens af den anden målnukleinsyresekvens, hvor den første målnukleinsyre eller den anden målnukleinsyre omfatter en T-cellereceptorsekvens eller en immunoglobulinsekvens; isolering af enkeltcellerne med mindst ét sæt af nukleinsyreprober; amplificering af den første og anden målnukleinsyresekvens uafhængigt, hvor den første målnukleinsyresekvens amplificeres ved anvendelse af den første probe og den anden probe, og hvor den anden målnukleinsyresekvens amplificeres ved anvendelse af den tredje probe og den fjerde probe; hybridisering af den eksogene sekvens til dens komplement; amplificering af den første målnukleinsyresekvens, den anden målnukleinsyresekvens og den eksogene sekvens ved anvendelse af den første og fjerde probe, til derved frembringelse af et fusioneret kompleks; og udførelse af en massesekventeringsreaktion til frembringelse af sekvensinformation for mindst 100.000 fusionerede komplekser fra mindst 10.000 celler i populationen af celler, hvor sekvensinformationen er tilstrækkelig til samlokalisering af den første målnukleinsyresekvens og den anden målnukleinsyresekvens til en enkeltcelle fra populationen af mindst 10.000 celler.
2. Fremgangsmåde ifølge krav 1, hvor amplificeringstrinet omfatter udførelse af (a) en polymerasekædereaktion, og hvor den første og tredje probe erfremad-primere, og den anden og fjerde probe er revers-primere til polymerasekædereaktionen; (b) en ligasekædereaktion; eller (c) en polymerasekædereaktion, en revers-transkriptase-polymerasekædereaktion eller en ligasekædereaktion efterfulgt af en polymerasekædereaktion.
3. Fremgangsmåde ifølge krav 1 eller 2, hvor det fusionerede kompleks er cirkulært.
4. Fremgangsmåde ifølge et af kravene 1-3, hvor den første eller anden målnukleinsyresekvens er en RNA-sekvens eller en DNA-sekvens.
5. Fremgangsmåde ifølge et af kravene 1-4, hvor (a) den første eller anden målnukleinsyresekvens omfatter en T-cellereceptorsekvens, hvor fortrinsvis den første målnukleinsyresekvens, den anden målnukleinsyresekvens eller begge målnukleinsyresekvenser omfatter en immunoglobulinsekvens; (b) den første målnukleinsyre omfatter en T-cellereceptorsekvens, og den anden målnukleinsyresekvens omfatter et andet molekyle, der er forbundet med immuncellefunktion; (c) den første målnukleinsyresekvens omfatter en immunoglobulinsekvens, og den anden sekvens omfatter et andet molekyle, der er forbundet med immuncellefunktion; eller (d) den første eller anden målnukleinsyre omfatter en sjælden gensekvens.
6. Fremgangsmåde ifølge krav 5(b), hvor det andet molekyle er udvalgt fra gruppen, der består af: interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-gamma (IFNy), interleukin-10 (IL-10), interleukin-1 (IL-1), interleukin-13 (IL-13), interleukin-17 (IL-17), interleukin-18 (IL-18), tumornekrosefaktor-alfa (TNFa), tumornekrosefaktor-beta (TNF3), T-box-transkriptionsfaktor 21 (TBX21), forkhead box P3 (FOXP3), differentieringsklynge 4 (CD4), differentieringsklynge 8 (CD8), differentieringsklynge 1d (CD1d), differentieringsklynge 161 (CD161), differentieringsklynge 3 (CD3), større histokompatibilitetskompleks (MHC), differentieringsklynge 19 (CD19), interleukin-7-receptor (IL-7-receptor), differentieringsklynge 10 (CD10), differentieringsklynge 20 (CD20), differentieringsklynge 22 (CD22), differentieringsklynge 34 (CD34), differentieringsklynge 27 (CD27), differentieringsklynge 5 (CD5) og differentieringsklynge 45 (CD45), differentieringsklynge 38 (CD38), differentieringsklynge 78 (CD78), interleukin-6-receptor, interferon-regulatorfaktor 4 (IRF4) og differentieringsklynge 138 (CD138).
7. Fremgangsmåde ifølge krav 5(c), hvor det andet molekyle er udvalgt fra gruppen, der består af: interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-gamma (IFNy), interleukin-10 (IL-10), interleukin-1 (IL-1), interleukin-13 (IL-13), interleukin-17 (IL-17), interleukin-18 (IL-18), tumornekrosefaktor-alfa (TNFa), tumornekrosefaktor-beta (TNF3), T-box-transkriptionsfaktor 21 (TBX21), forkhead box P3 (FOXP3), differentieringsklynge 4 (CD4), differentieringsklynge 8 (CD8), differentieringsklynge 1d (CD1d), differentieringsklynge 161 (CD161), differentieringsklynge 3 (CD3), større histokompatibilitetskompleks (MHC), differentieringsklynge 19 (CD19), interleukin-7-receptor (IL-7-receptor), differentieringsklynge 10 (CD10), differentieringsklynge 20 (CD20), differentieringsklynge 22 (CD22), differentieringsklynge 34 (CD34), differentieringsklynge 27 (CD27), differentieringsklynge 5 (CD5) og differentieringsklynge 45 (CD45), differentieringsklynge 38 (CD38), differentieringsklynge 78 (CD78), interleukin-6-receptor, interferon-regulatorfaktor 4 (IRF4) og differentieringsklynge 138 (CD138).
8. Fremgangsmåde ifølge krav 5(d), hvor (a) den sjældne gensekvens er til stede i færre end 5 % af cellerne, fortrinsvis i færre end 1 % af cellerne og mere fortrinsvis i færre end 0,1 % af cellerne; eller (b) den sjældne gensekvens er resultatet af en genmutation, hvor fortrinsvis (ba) mutationen er en somatisk mutation; (bb) mutationen er forbundet med en sygdom; eller (bc) genmutationen er en mutation i et gen udvalgt fra gruppen, der består af epidermal vækstfaktorreceptor (EGFR), phosphatase og tensinhomolog (PTEN), tumorprotein 53 (p53), MutS-homolog 2 (MSH2), multipel endokrin neoplasi 1 (MEN1), adenomatøs polyposis coli (APC), Fas-receptor (FASR), retinoblastomprotein (Rb1), Janus-kinase 2 (JAK2), (ETS)-lignende transkriptionsfaktor 1 (ELK1), v-ets aviær erytroblastosevirus E26-onkogenhomolog 1 (ETS1), brystcancer 1 (BRCA1), brystcancer 2 (BRCA2), hepatocyt-vækstfaktorreceptor (MET), ret proto-onkogen (RET), V-erb-b2 erytroblastisk leukæmi viral onkogenhomolog 2 (HER2), V-Ki-ras2 Kirsten rottesarkom viral onkogenhomolog (KRAS), B-cellelymfom 2 (BCL2), V-myc myelocytomatose viral onkogenhomolog (MYC), neurofibromatose type 2-gen (NF2), v-myb myeloblastose viral onkogenhomolog (MYB) og mutS-homolog 6 (E. coli) (MSH6); hvor sygdommen fortrinsvis er cancer, og canceren fortrinsvis er udvalgt fra gruppen, der består af: lungekarcinom, ikke-småcellet lungecancer, småcellet lungecancer, uteruscancer, thyreoideacancer, brystkarcinom, prostatakarcinom, pancreaskarcinom, colonkarcinom, lymfom, Burkitts lymfom, Hodgkins lymfom, myeloid leukæmi, leukæmi, sarkom, biastom, melanom, seminom, hjernecancer, gliom, glioblastom, cerebellar astrocytom, kutan T-cellelymform, gastrisk cancer, levercancer, ependymom, larynxcancer, halscancer, mavecancer, nyrecancer, pancreascancer, blærecancer, øsofaguscancer, testikelcancer, medulloblastom, vaginalcancer, ovariecancer, cervixcancer, basalcellekarcinom, hypofyseadenom, rabdomyosarkom og Kaposis sarkom.
9. Fremgangsmåde ifølge et af kravene 1-8, der yderligere omfatter fiksering og permeabilisering af cellerne før udførelse af amplifikationstrinet.
10. Fremgangsmåde ifølge et af kravene 1-9, der yderligere omfatter lysering af cellerne før udførelse af amplifikationstrinet.
11. Fremgangsmåde ifølge et af kravene 1-10, der yderligere omfatter kvantificering af sekvensinformationen frembragt ved massesekventeringsreaktionen.
12. Fremgangsmåde ifølge et af kravene 1-11, hvor udførelse af massesekventeringsreaktionen til frembringelse af sekvensinformation udføres for mindst 1.000.000 fusionerede komplekser fra mindst 10.000 celler i populationen af celler.
13. Fremgangsmåde ifølge krav 1, der yderligere omfatter: (A) tilvejebringelse af et andet sæt af nukleinsyreprober, hvilket andet sæt omfatter en femte probe, der omfatter en sekvens, der er komplementær til en tredje målnukleinsyreundersekvens, en sjette probe, der omfatter en sekvens, der er komplementærtil en anden undersekvens af den tredje målnukleinsyresekvens, og en anden sekvens, der er komplementær til en anden eksogen sekvens, en syvende probe, der omfatter den anden eksogene sekvens og en sekvens, der er komplementærtil en første undersekvens af en fjerde målnukleinsyresekvens, og en ottende probe, der omfatter en sekvens, der er komplementærtil en anden undersekvens af den fjerde målnukleinsyresekvens; isolering af enkeltcellerne med det første og andet sæt af nukleinsyreprober; amplificering af den tredje og fjerde målnukleinsyresekvens uafhængigt, hvor den tredje målnukleinsyresekvens amplificeres ved anvendelse af den femte probe og den sjette probe, og hvor den fjerde målnukleinsyresekvens amplificeres ved anvendelse af den syvende probe og den ottende probe; hybridisering af den anden eksogene sekvens til dens komplement; amplificering af den tredje målnukleinsyresekvens, den fjerde målnukleinsyresekvens og den anden eksogene sekvens ved anvendelse af den femte og ottende probe til derved frembringelse af et fusioneret kompleks; og udførelse af en massesekventeringsreaktion til frembringelse af sekvensinformation for mindst 100.000 fusionerede komplekser fra mindst 10.000 celler i populationen af celler, hvor sekvensinformationen er tilstrækkelig til samlokalisering af den første målnukleinsyresekvens, den anden målnukleinsyresekvens, den tredje målnukleinsyresekvens og den fjerde målnukleinsyresekvens til en enkeltcelle fra populationen af mindst 10.000 celler; eller (B) tilvejebringelse af N sæt af nukleinsyreprober, hvor hver af de N sæt omfatter en lr probe, der omfatter en sekvens, der er komplementær til en la-målnukleinsyre-første undersekvens, en l2-probe, der omfatter en sekvens, der er komplementær til en la- målnukleinsyre-anden undersekvens, og en anden sekvens, der er komplementær til en eksogen l-sekvens, en b-probe, der omfatter den eksogene l-sekvens og en sekvens, der er komplementær til en lb-målnukleinsyre-første undersekvens, og en U- probe, der omfatter en sekvens, der er komplementær til en lb-målnukleinsyre-anden undersekvens, hvor I er i intervallet fra 1 til N; isolering af enkeltcellerne med de N sæt af nukleinsyreprober; amplificering for alle værdier af I af la- og lb-målnukleinsyresekvenserne uafhængigt, hvor la-målnukleinsyresekvensen amplificeres ved anvendelse af h-proben og b- proben, og lb-målnukleinsyresekvensen amplificeres ved anvendelse af b-proben og U-proben; hybridisering af den eksogene l-sekvens til dens komplement; amplificering for hver I af la-målsekvensen, b-målsekvensen og den eksogene I-sekvens ved anvendelse af h- og U-probeme til derved frembringelse af N fusionerede komplekser; og udførelse af en massesekventeringsreaktion til frembringelse af sekvensinformation for mindst 100.000 fusionerede komplekser fra mindst 10.000 celler i populationen af celler, hvor sekvensinformationen er tilstrækkelig til samlokalisering af den N la-målnukleinsyresekvens og lb-målnukleinsyresekvens til en enkeltcelle fra populationen af mindst 10.000 celler.
14. Fremgangsmåde til analyse af mindst to nukleinsyresekvenser i en enkeltcelle, der er indeholdt i en population af mindst 10.000 celler, som omfatter: isolering af hver af et antal enkeltceller fra en population af mindst 10.000 celler i en emulsionsmikrodråbe, indførelse af en unik eksogen stregkodesekvens, der omfatter mindst seks nukleotider, i hver af emulsionsmikrodråberne, hvor hver stregkodesekvens er udvalgt fra en pulje af stregkodesekvenser med mere end 1000-fold diversitet i sekvens, hvor hver af de unikke eksogene stregkodesekvenser matches med en enkeltcelle fra antallet af enkeltceller; for hver af antallet af enkeltceller tilvejebringelse af mindst ét sæt af nukleinsyreprober, hvilket sæt omfatter en første probe, der omfatter en sekvens, der er komplementærtil en nukleinsyresekvens, der er placeret i 5-enden af den unikke eksogene stregkodesekvens, en anden probe, der omfatter en sekvens, der er komplementær til en nukleinsyresekvens der er placeret i 3-enden af den unikke eksogene stregkodesekvens, og en anden region af sekvens, der er komplementær til en ikke-human, eksogene sekvens, en tredje probe, der omfatter en sekvens, der omfatter den ikke-humane, eksogene sekvens, og en sekvens, der er komplementær til en første undersekvens af en anden målnukleinsyresekvens, og en fjerde probe, der omfatter en sekvens, der er komplementærtil en anden undersekvens af den anden målnukleinsyresekvens, og hvor den anden målnukleinsyresekvens omfatter en T-cellereceptorsekvens eller en immunoglobulinsekvens; amplificering af den første og anden nukleinsyresekvens uafhængigt, hvor den eksogene stregkodesekvens amplificeres ved anvendelse af den første probe og den anden probe, og hvor den anden målnukleinsyresekvens amplificeres ved anvendelse af den tredje probe og den fjerde probe; hybridisering af den ikke-humane, eksogene sekvens til dens komplement; amplificering af stregkodesekvensen, den anden målnukleinsyresekvens og den ikke-humane, eksogene sekvens ved anvendelse af den første og fjerde probe; udførelse af massesekventering af de fusionerede komplekser; og identificering af en enkeltcelle for hver af de fusionerede komplekser på basis af stregkodesekvensen.
15. Fremgangsmåde ifølge krav 14, hvor stregkodesekvensen fikseres til et korn eller en fast overflade, hvor fortrinsvis kornet eller den faste overflade isoleres i emulsionsmikrodråben eller reaktionsbeholderen, hvor fortrinsvis indførelse afen unik stregkodesekvens omfatter fusionering af emulsionsmikrodråben eller en reaktionsbeholder, der omfatter enkeltcellen, med emulsionsmikrodråben eller en reaktionsbeholder, der omfatter stregkodesekvensen, der erfikseret til kornet eller den faste overflade.
16. Fremgangsmåde ifølge krav 14 eller krav 15, hvor den anden målnukleinsyresekvens er komplementær til en RNA-sekvens eller komplementær til en DNA-sekvens.
17. Fremgangsmåde ifølge et af kravene 14-16, hvor amplificering omfatter (a) udførelse af en polymerasekædereaktion; (b) udførelse af en ligasekædereaktion; eller (c) udførelse af en ligasekædereaktion efterfulgt af polymerasekædereaktion.
18. Fremgangsmåde ifølge krav 1 eller krav 14, hvor enkeltcellen er indeholdt i en population af mindst 25.000 celler, mindst 50.000 celler, mindst 75.000 celler eller mindst 100.000 celler.
19. Fremgangsmåde ifølge krav 14, der yderligere omfatter kvantificering af de fusionerede komplekser.
20. Fremgangsmåde ifølge et af kravene 14-19, hvor de fusionerede komplekser er cirkulære.
21. Fremgangsmåde ifølge krav 14, der yderligere omfatter: tilvejebringelse af N sæt af nukleinsyreprober, hvor hvert af de N sæt omfatter en h-probe, der omfatter en sekvens, der er komplementær til en første undersekvens af en stregkodesekvens, en b-probe, der omfatter en sekvens, der er komplementær til en anden undersekvens af stregkodesekvensen, og en anden sekvens, der er komplementærtil en eksogen l-sekvens, en h-probe, der omfatter den eksogene I-sekvens og en sekvens, der er komplementærtil en lb-målnukleinsyre-første undersekvens, og en U-probe, der omfatter en sekvens, der er komplementær til en lb-målnukleinsyre-anden undersekvens, hvori er i intervallet fra 1 til N; isolering af enkeltcellerne med de N sæt af nukleinsyreprober; amplificering for alle værdier af I af stregkodesekvensen og lb-målnukleinsyresekvenserne uafhængigt, hvor stregkodesekvensen amplificeres ved anvendelse af h-proben og h-proben, og lb-målnukleinsyresekvensen amplificeres ved anvendelse af h-proben og U-proben; hybridisering af den eksogene l-sekvens til dens komplement; amplificering for hver I af stregkodesekvensen, lb-målsekvensen og den eksogene I-sekvens ved anvendelse af h- og U-proben til derved frembringelse af N fusionerede komplekser; og udførelse af en massesekventeringsreaktion til frembringelse af sekvensinformation for mindst 100.000 fusionerede komplekser fra mindst 10.000 celler i populationen af celler, hvor sekvensinformationen er tilstrækkelig til samlokalisering af stregkodesekvensen og lb-målnukleinsyresekvensen til en enkeltcelle fra populationen af mindst 10.000 celler.
22. Fremgangsmåde ifølge krav 13(B) eller krav 21, hvor N er mindre end eller lig med 10, mindre end eller lig med 100, mindre end eller lig med 1000, mindre end eller lig med 10.000, mindre end eller lig med 100.000, eller N er alle de polyadenylerede transkripter i en celle.
23. Fremgangsmåde ifølge krav 21 eller krav 22, hvor stregkodesekvensen er den samme sekvens for alle N.
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Families Citing this family (271)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7968287B2 (en) 2004-10-08 2011-06-28 Medical Research Council Harvard University In vitro evolution in microfluidic systems
EP2364774A3 (en) 2006-01-11 2014-06-04 Raindance Technologies, Inc. Microfluidic Devices And Methods Of Use In The Formation And Control Of Nanoreactors
ATE540750T1 (de) 2006-05-11 2012-01-15 Raindance Technologies Inc Mikrofluidische vorrichtung und verfahren
US9562837B2 (en) 2006-05-11 2017-02-07 Raindance Technologies, Inc. Systems for handling microfludic droplets
US8772046B2 (en) 2007-02-06 2014-07-08 Brandeis University Manipulation of fluids and reactions in microfluidic systems
WO2008130623A1 (en) 2007-04-19 2008-10-30 Brandeis University Manipulation of fluids, fluid components and reactions in microfluidic systems
EP4047367A1 (en) 2008-07-18 2022-08-24 Bio-Rad Laboratories, Inc. Method for detecting target analytes with droplet libraries
US12038438B2 (en) 2008-07-18 2024-07-16 Bio-Rad Laboratories, Inc. Enzyme quantification
US8748103B2 (en) 2008-11-07 2014-06-10 Sequenta, Inc. Monitoring health and disease status using clonotype profiles
US9365901B2 (en) 2008-11-07 2016-06-14 Adaptive Biotechnologies Corp. Monitoring immunoglobulin heavy chain evolution in B-cell acute lymphoblastic leukemia
US8628927B2 (en) 2008-11-07 2014-01-14 Sequenta, Inc. Monitoring health and disease status using clonotype profiles
US9528160B2 (en) 2008-11-07 2016-12-27 Adaptive Biotechnolgies Corp. Rare clonotypes and uses thereof
WO2010053587A2 (en) 2008-11-07 2010-05-14 Mlc Dx Incorporated Methods of monitoring conditions by sequence analysis
US9506119B2 (en) 2008-11-07 2016-11-29 Adaptive Biotechnologies Corp. Method of sequence determination using sequence tags
US9394567B2 (en) 2008-11-07 2016-07-19 Adaptive Biotechnologies Corporation Detection and quantification of sample contamination in immune repertoire analysis
PL2387627T3 (pl) 2009-01-15 2016-10-31 Profilowanie odporności adaptacyjnej i wytwarzanie przeciwciał monoklonalnych
US8528589B2 (en) 2009-03-23 2013-09-10 Raindance Technologies, Inc. Manipulation of microfluidic droplets
KR20120044941A (ko) 2009-06-25 2012-05-08 프레드 헛친슨 켄서 리서치 센터 적응 면역의 측정방법
US9315857B2 (en) 2009-12-15 2016-04-19 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse label-tags
US8835358B2 (en) 2009-12-15 2014-09-16 Cellular Research, Inc. Digital counting of individual molecules by stochastic attachment of diverse labels
US10351905B2 (en) * 2010-02-12 2019-07-16 Bio-Rad Laboratories, Inc. Digital analyte analysis
US9399797B2 (en) 2010-02-12 2016-07-26 Raindance Technologies, Inc. Digital analyte analysis
EP3392349A1 (en) 2010-02-12 2018-10-24 Raindance Technologies, Inc. Digital analyte analysis
US20190300945A1 (en) 2010-04-05 2019-10-03 Prognosys Biosciences, Inc. Spatially Encoded Biological Assays
US10787701B2 (en) 2010-04-05 2020-09-29 Prognosys Biosciences, Inc. Spatially encoded biological assays
EP2622103B2 (en) 2010-09-30 2022-11-16 Bio-Rad Laboratories, Inc. Sandwich assays in droplets
DK2652155T3 (da) 2010-12-16 2017-02-13 Gigagen Inc Fremgangsmåder til massiv parallelanalyse af nukleinsyrer i enkeltceller
US10144950B2 (en) 2011-01-31 2018-12-04 Roche Sequencing Solutions, Inc. Methods of identifying multiple epitopes in cells
EP3859011A1 (en) 2011-02-11 2021-08-04 Bio-Rad Laboratories, Inc. Methods for forming mixed droplets
EP2675819B1 (en) 2011-02-18 2020-04-08 Bio-Rad Laboratories, Inc. Compositions and methods for molecular labeling
WO2012129363A2 (en) * 2011-03-24 2012-09-27 President And Fellows Of Harvard College Single cell nucleic acid detection and analysis
GB201106254D0 (en) 2011-04-13 2011-05-25 Frisen Jonas Method and product
EP3709018A1 (en) 2011-06-02 2020-09-16 Bio-Rad Laboratories, Inc. Microfluidic apparatus for identifying components of a chemical reaction
US8658430B2 (en) 2011-07-20 2014-02-25 Raindance Technologies, Inc. Manipulating droplet size
US10385475B2 (en) 2011-09-12 2019-08-20 Adaptive Biotechnologies Corp. Random array sequencing of low-complexity libraries
CA2853088C (en) 2011-10-21 2018-03-13 Adaptive Biotechnologies Corporation Quantification of adaptive immune cell genomes in a complex mixture of cells
EP3904536B1 (en) 2011-12-09 2025-10-29 Adaptive Biotechnologies Corporation Diagnosis of lymphoid malignancies and minimal residual disease detection
US9499865B2 (en) 2011-12-13 2016-11-22 Adaptive Biotechnologies Corp. Detection and measurement of tissue-infiltrating lymphocytes
EP3309262B1 (en) * 2012-02-24 2019-09-25 Bio-Rad Laboratories, Inc. Labeling and sample preparation for sequencing
WO2013130512A2 (en) 2012-02-27 2013-09-06 The University Of North Carolina At Chapel Hill Methods and uses for molecular tags
ES2904816T3 (es) 2012-02-27 2022-04-06 Becton Dickinson Co Composiciones para recuento molecular
WO2013134261A1 (en) 2012-03-05 2013-09-12 President And Fellows Of Harvard College Systems and methods for epigenetic sequencing
US10077478B2 (en) 2012-03-05 2018-09-18 Adaptive Biotechnologies Corp. Determining paired immune receptor chains from frequency matched subunits
EP3744857A1 (en) * 2012-03-20 2020-12-02 University Of Washington Through Its Center For Commercialization Methods of lowering the error rate of massively parallel dna sequencing using duplex consensus sequencing
JP2015512655A (ja) * 2012-04-13 2015-04-30 シーケンタ インコーポレイテッド 免疫レパートリ分析におけるサンプル混入の検出および定量化
KR101621643B1 (ko) 2012-05-08 2016-05-16 어댑티브 바이오테크놀로지스 코포레이션 다중 pcr 반응에서 증폭 바이어스를 측정 및 교정하기 위한 조성물 및 방법
ES2774165T3 (es) 2012-06-15 2020-07-17 Univ Texas Secuenciación de alto rendimiento de múltiples transcritos de una única célula
EP2861761A1 (en) 2012-06-15 2015-04-22 Adaptive Biotechnologies Corporation Uniquely tagged rearranged adaptive immune receptor genes in a complex gene set
WO2014026032A2 (en) 2012-08-08 2014-02-13 Apprise Bio, Inc. Increasing dynamic range for identifying multiple epitopes in cells
US10161007B2 (en) 2012-08-13 2018-12-25 The Regents Of The University Of California Methods and systems for detecting biological components
US10584381B2 (en) 2012-08-14 2020-03-10 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
US10221442B2 (en) 2012-08-14 2019-03-05 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
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
US10273541B2 (en) 2012-08-14 2019-04-30 10X Genomics, Inc. Methods and systems for processing polynucleotides
US11591637B2 (en) 2012-08-14 2023-02-28 10X Genomics, Inc. Compositions and methods for sample processing
BR112015003354A8 (pt) 2012-08-14 2018-01-16 10X Genomics Inc métodos e composições de microcápsula
EP2898096B1 (en) 2012-09-21 2024-02-14 The Broad Institute, Inc. Methods for labeling of rnas
EP3640343B1 (en) 2012-10-01 2026-01-07 Adaptive Biotechnologies Corporation Immunocompetence assessment by adaptive immune receptor diversity and clonality characterization
EP3901280B1 (en) 2012-10-17 2025-03-12 10x Genomics Sweden AB Methods and product for optimising localised or spatial detection of gene expression in a tissue sample
WO2015160439A2 (en) 2014-04-17 2015-10-22 Adaptive Biotechnologies Corporation Quantification of adaptive immune cell genomes in a complex mixture of cells
AU2013359165B2 (en) 2012-12-14 2019-09-12 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
WO2014116729A2 (en) * 2013-01-22 2014-07-31 The Board Of Trustees Of The Leland Stanford Junior University Haplotying of hla loci with ultra-deep shotgun sequencing
CA2900543C (en) 2013-02-08 2023-01-31 10X Genomics, Inc. Partitioning and processing of analytes and other species
EP3647436A1 (en) * 2013-03-14 2020-05-06 Quest Diagnostics Investments Incorporated Method for detecting cystic fibrosis
US9816088B2 (en) 2013-03-15 2017-11-14 Abvitro Llc Single cell bar-coding for antibody discovery
CA2906218A1 (en) 2013-03-15 2014-09-18 Adaptive Biotechnologies Corporation Uniquely tagged rearranged adaptive immune receptor genes in a complex gene set
CN105473744B (zh) 2013-06-25 2020-06-19 普罗格诺西斯生物科学公司 采用微流控装置的空间编码生物分析
US9708657B2 (en) 2013-07-01 2017-07-18 Adaptive Biotechnologies Corp. Method for generating clonotype profiles using sequence tags
EP3480321B8 (en) 2013-08-28 2021-03-10 Becton, Dickinson and Company Massively parallel single cell analysis
US10395758B2 (en) 2013-08-30 2019-08-27 10X Genomics, Inc. Sequencing methods
US11901041B2 (en) 2013-10-04 2024-02-13 Bio-Rad Laboratories, Inc. Digital analysis of nucleic acid modification
CN105745528A (zh) 2013-10-07 2016-07-06 赛卢拉研究公司 用于以数字方式对阵列上的特征进行计数的方法和系统
US10655173B2 (en) 2013-10-18 2020-05-19 The Broad Institute, Inc. Spatial and cellular mapping of biomolecules in situ by high-throughput sequencing
US9944977B2 (en) 2013-12-12 2018-04-17 Raindance Technologies, Inc. Distinguishing rare variations in a nucleic acid sequence from a sample
US9824068B2 (en) 2013-12-16 2017-11-21 10X Genomics, Inc. Methods and apparatus for sorting data
SG11201605344YA (en) 2013-12-30 2016-07-28 Atreca Inc Analysis of nucleic acids associated with single cells using nucleic acid barcodes
US10537889B2 (en) * 2013-12-31 2020-01-21 Illumina, Inc. Addressable flow cell using patterned electrodes
ES2741740T3 (es) 2014-03-05 2020-02-12 Adaptive Biotechnologies Corp Métodos que usan moléculas sintéticas que contienen segmentos de nucleótidos aleatorios
US10066265B2 (en) 2014-04-01 2018-09-04 Adaptive Biotechnologies Corp. Determining antigen-specific t-cells
DE202015009609U1 (de) 2014-04-10 2018-08-06 10X Genomics, Inc. Mikrofluidisches System zur Erzeugung von Emulsionen
US20150298091A1 (en) 2014-04-21 2015-10-22 President And Fellows Of Harvard College Systems and methods for barcoding nucleic acids
LT3299469T (lt) * 2014-04-21 2020-04-27 President And Fellows Of Harvard College Sistemos ir būdai, skirti nukleorūgšties identifikavimo žymės įvedimui
US10202640B2 (en) 2014-05-07 2019-02-12 The Board Of Trustees Of The Leland Stanford Junior University Single cell analysis of T cells using high-throughput multiplex amplification and deep sequencing
AU2015279546B2 (en) 2014-06-26 2021-04-08 10X Genomics, Inc. Processes and systems for nucleic acid sequence assembly
WO2015200893A2 (en) 2014-06-26 2015-12-30 10X Genomics, Inc. Methods of analyzing nucleic acids from individual cells or cell populations
US12312640B2 (en) 2014-06-26 2025-05-27 10X Genomics, Inc. Analysis of nucleic acid sequences
CN106536756A (zh) 2014-06-26 2017-03-22 10X基因组学有限公司 核酸序列的分析
EP3160654A4 (en) 2014-06-27 2017-11-15 The Regents of The University of California Pcr-activated sorting (pas)
US10526641B2 (en) * 2014-08-01 2020-01-07 Dovetail Genomics, Llc Tagging nucleic acids for sequence assembly
CN107002076B (zh) * 2014-09-15 2021-11-23 阿布维特罗有限责任公司 高通量核苷酸文库测序
US10434507B2 (en) 2014-10-22 2019-10-08 The Regents Of The University Of California High definition microdroplet printer
EP3715455A1 (en) 2014-10-29 2020-09-30 Adaptive Biotechnologies Corp. Highly-multiplexed simultaneous detection of nucleic acids encoding paired adaptive immune receptor heterodimers from many samples
EP3212807B1 (en) 2014-10-29 2020-09-02 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
US10246701B2 (en) 2014-11-14 2019-04-02 Adaptive Biotechnologies Corp. Multiplexed digital quantitation of rearranged lymphoid receptors in a complex mixture
WO2016077750A1 (en) 2014-11-14 2016-05-19 Athena Diagnostics, Inc. Methods to detect a silent carrier genotype
US10900065B2 (en) * 2014-11-14 2021-01-26 University Of Washington Methods and kits for labeling cellular molecules
WO2016086029A1 (en) 2014-11-25 2016-06-02 Adaptive Biotechnologies Corporation Characterization of adaptive immune response to vaccination or infection using immune repertoire sequencing
DK3227684T3 (da) 2014-12-03 2019-11-11 Isoplexis Corp Analyse af og screening for cellesekretionsprofiler
AU2016207023B2 (en) 2015-01-12 2019-12-05 10X Genomics, Inc. Processes and systems for preparing nucleic acid sequencing libraries and libraries prepared using same
WO2016115273A1 (en) 2015-01-13 2016-07-21 10X Genomics, Inc. Systems and methods for visualizing structural variation and phasing information
CN104531713A (zh) * 2015-01-20 2015-04-22 中国人民解放军第三军医大学 基于高通量测序构建人tcbr文库的多重pcr引物和方法
CA2974306A1 (en) 2015-02-04 2016-08-11 The Regents Of The University Of California Sequencing of nucleic acids via barcoding in discrete entities
AU2016219480B2 (en) 2015-02-09 2021-11-11 10X Genomics, Inc. Systems and methods for determining structural variation and phasing using variant call data
US10697010B2 (en) 2015-02-19 2020-06-30 Becton, Dickinson And Company High-throughput single-cell analysis combining proteomic and genomic information
EP3262407B1 (en) 2015-02-24 2023-08-30 10X Genomics, Inc. Partition processing methods and systems
CN115651972A (zh) 2015-02-24 2023-01-31 10X 基因组学有限公司 用于靶向核酸序列覆盖的方法
ES3002665T3 (en) 2015-02-24 2025-03-07 Adaptive Biotechnologies Corp Methods for diagnosing infectious disease and determining hla status using immune repertoire sequencing
ES2836802T3 (es) 2015-02-27 2021-06-28 Becton Dickinson Co Códigos de barras moleculares espacialmente direccionables
CN107614096A (zh) * 2015-03-13 2018-01-19 哈佛学院院长及董事 使用扩增测定细胞
US10513733B2 (en) 2015-03-23 2019-12-24 Board Of Regents, The University Of Texas System High throughout sequencing of paired VH and VL transcripts from B cells secreting antigen-specific antibodies
WO2016154540A1 (en) 2015-03-26 2016-09-29 Dovetail Genomics Llc Physical linkage preservation in dna storage
EP4180535A1 (en) 2015-03-30 2023-05-17 Becton, Dickinson and Company Methods and compositions for combinatorial barcoding
AU2016242967B2 (en) 2015-04-01 2021-07-01 Adaptive Biotechnologies Corp. Method of identifying human compatible T cell receptors specific for an antigenic target
FI3901281T3 (fi) 2015-04-10 2023-01-31 Biologisten näytteiden spatiaalisesti eroteltu moninkertainen nukleiinihappoanalyysi
US11746367B2 (en) 2015-04-17 2023-09-05 President And Fellows Of Harvard College Barcoding systems and methods for gene sequencing and other applications
CN107580632B (zh) 2015-04-23 2021-12-28 贝克顿迪金森公司 用于全转录组扩增的方法和组合物
WO2016196229A1 (en) 2015-06-01 2016-12-08 Cellular Research, Inc. Methods for rna quantification
US9422547B1 (en) 2015-06-09 2016-08-23 Gigagen, Inc. Recombinant fusion proteins and libraries from immune cell repertoires
DK3307905T3 (da) 2015-06-09 2022-02-14 Gigagen Inc Rekombinante fusionsproteiner og biblioteker fra immuncellerepertoirer
JP6940484B2 (ja) 2015-09-11 2021-09-29 セルラー リサーチ， インコーポレイテッド ライブラリー正規化のための方法および組成物
JP7064439B2 (ja) * 2015-11-04 2022-05-10 アトレカ インコーポレイテッド 単一細胞に関連する核酸の解析のための、核酸バーコードの組み合わせセット
US11371094B2 (en) 2015-11-19 2022-06-28 10X Genomics, Inc. Systems and methods for nucleic acid processing using degenerate nucleotides
DK3882357T3 (da) 2015-12-04 2022-08-29 10X Genomics Inc Fremgangsmåder og sammensætninger til analyse af nukleinsyrer
EP3387152B1 (en) 2015-12-08 2022-01-26 Twinstrand Biosciences, Inc. Improved adapters, methods, and compositions for duplex 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
CA3014911A1 (en) 2016-02-23 2017-08-31 Dovetail Genomics, Llc Generation of phased read-sets for genome assembly and haplotype phasing
EP3443087B1 (en) * 2016-04-12 2024-09-18 Medimmune, LLC Immune repertoire mining
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
EP4269616A3 (en) 2016-05-02 2024-02-14 Becton, Dickinson and Company Accurate molecular barcoding
WO2017197343A2 (en) 2016-05-12 2017-11-16 10X Genomics, Inc. Microfluidic on-chip filters
DK3455356T3 (da) 2016-05-13 2021-11-01 Dovetail Genomics Llc Genfinding af langtrækkende bindingsinformation fra konserverede prøver
WO2017197338A1 (en) 2016-05-13 2017-11-16 10X Genomics, Inc. Microfluidic systems and methods of use
US10301677B2 (en) 2016-05-25 2019-05-28 Cellular Research, Inc. Normalization of nucleic acid libraries
CN109074430B (zh) 2016-05-26 2022-03-29 贝克顿迪金森公司 分子标记计数调整方法
US10640763B2 (en) 2016-05-31 2020-05-05 Cellular Research, Inc. Molecular indexing of internal sequences
US10202641B2 (en) 2016-05-31 2019-02-12 Cellular Research, Inc. Error correction in amplification of samples
EP3686287A1 (en) * 2016-06-28 2020-07-29 Hifibio Method for transcriptome analysis of single cells
US10465242B2 (en) 2016-07-14 2019-11-05 University Of Utah Research Foundation Multi-sequence capture system
CN110088290A (zh) 2016-08-10 2019-08-02 加利福尼亚大学董事会 在乳液微滴中结合多重置换扩增和pcr
CN109964126B (zh) 2016-09-12 2022-12-27 伊索普莱克西斯公司 用于细胞治疗法和其他免疫治疗法的多重分析的系统和方法
US10428325B1 (en) 2016-09-21 2019-10-01 Adaptive Biotechnologies Corporation Identification of antigen-specific B cell receptors
CN109791157B (zh) 2016-09-26 2022-06-07 贝克顿迪金森公司 使用具有条形码化的寡核苷酸序列的试剂测量蛋白质表达
EP3529400B1 (en) 2016-10-24 2021-02-17 Geneinfosec, Inc. Concealing information present within nucleic acids
JP7228510B2 (ja) 2016-11-08 2023-02-24 ベクトン・ディキンソン・アンド・カンパニー 細胞標識分類の方法
ES2980967T3 (es) 2016-11-08 2024-10-03 Becton Dickinson Co Métodos para la clasificación de perfiles de expresión
DK3538891T3 (da) * 2016-11-11 2022-03-28 Isoplexis Corp Sammensætninger og fremgangsmåder til samtidig genomisk, transkriptomisk og proteomisk analyse af enkeltceller
EP3559231B1 (en) * 2016-11-11 2022-08-17 Bio-Rad Laboratories, Inc. Methods for processing nucleic acid samples
US11525783B2 (en) 2016-11-22 2022-12-13 IsoPlexis Corporation Systems, devices and methods for cell capture and methods of manufacture thereof
AU2017382905A1 (en) 2016-12-21 2019-07-04 The Regents Of The University Of California Single cell genomic sequencing using hydrogel based droplets
US10011872B1 (en) 2016-12-22 2018-07-03 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
US10815525B2 (en) 2016-12-22 2020-10-27 10X Genomics, Inc. Methods and systems for processing polynucleotides
CN110366557B (zh) 2016-12-23 2024-04-09 威特拉公司 结合多肽及其制备方法
JP6234542B1 (ja) 2016-12-27 2017-11-22 株式会社 TL Genomics 胎児細胞由来染色体ｄｎａの取得方法
ES2961580T3 (es) 2017-01-13 2024-03-12 Cellular Res Inc Revestimiento hidrófilo de canales de fluidos
US12264411B2 (en) 2017-01-30 2025-04-01 10X Genomics, Inc. Methods and systems for analysis
EP4029939B1 (en) 2017-01-30 2023-06-28 10X Genomics, Inc. Methods and systems for droplet-based single cell barcoding
CN110382708A (zh) 2017-02-01 2019-10-25 赛卢拉研究公司 使用阻断性寡核苷酸进行选择性扩增
US10995333B2 (en) 2017-02-06 2021-05-04 10X Genomics, Inc. Systems and methods for nucleic acid preparation
CN110431233B (zh) * 2017-03-13 2022-08-12 吉加根公司 用于单细胞的大量平行组合分析的系统和方法
CN110870018B (zh) 2017-05-19 2024-11-22 10X基因组学有限公司 用于分析数据集的系统和方法
CN116064732A (zh) 2017-05-26 2023-05-05 10X基因组学有限公司 转座酶可接近性染色质的单细胞分析
US10400235B2 (en) 2017-05-26 2019-09-03 10X Genomics, Inc. Single cell analysis of transposase accessible chromatin
KR102790050B1 (ko) 2017-06-05 2025-04-04 백톤 디킨슨 앤드 컴퍼니 단일 세포를 위한 샘플 인덱싱
US10837047B2 (en) 2017-10-04 2020-11-17 10X Genomics, Inc. Compositions, methods, and systems for bead formation using improved polymers
US10501739B2 (en) 2017-10-18 2019-12-10 Mission Bio, Inc. Method, systems and apparatus for single cell analysis
WO2019084043A1 (en) 2017-10-26 2019-05-02 10X Genomics, Inc. METHODS AND SYSTEMS FOR NUCLEIC ACID PREPARATION AND CHROMATIN ANALYSIS
US11866785B2 (en) 2017-10-27 2024-01-09 Board Of Regents, The University Of Texas System Tumor specific antibodies and T-cell receptors and methods of identifying the same
CN111479631B (zh) 2017-10-27 2022-02-22 10X基因组学有限公司 用于样品制备和分析的方法和系统
ES2977793T3 (es) 2017-11-08 2024-08-30 Twinstrand Biosciences Inc Reactivos y adaptadores para la secuenciación de ácidos nucleicos y métodos para fabricar dichos reactivos y adaptadores
WO2019099751A1 (en) 2017-11-15 2019-05-23 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
US11254980B1 (en) 2017-11-29 2022-02-22 Adaptive Biotechnologies Corporation Methods of profiling targeted polynucleotides while mitigating sequencing depth requirements
WO2019108851A1 (en) 2017-11-30 2019-06-06 10X Genomics, Inc. Systems and methods for nucleic acid preparation and analysis
WO2019118355A1 (en) 2017-12-12 2019-06-20 10X Genomics, Inc. Systems and methods for single cell processing
CN111492068B (zh) 2017-12-19 2025-03-21 贝克顿迪金森公司 与寡核苷酸相关联的颗粒
CN118547046A (zh) 2017-12-22 2024-08-27 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
CN112005115A (zh) 2018-02-12 2020-11-27 10X基因组学有限公司 表征来自单个细胞或细胞群体的多种分析物的方法
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
EP3775271B1 (en) 2018-04-06 2025-03-12 10X Genomics, Inc. Systems and methods for quality control in single cell processing
EP3788170B1 (en) 2018-05-03 2025-01-01 Becton, Dickinson and Company Molecular barcoding on opposite transcript ends
US11773441B2 (en) 2018-05-03 2023-10-03 Becton, Dickinson And Company High throughput multiomics sample analysis
US11649482B2 (en) * 2018-05-08 2023-05-16 President And Fellows Of Harvard College Droplet-based single cell genomic DNA sequencing
WO2019217758A1 (en) 2018-05-10 2019-11-14 10X Genomics, Inc. Methods and systems for molecular library generation
WO2019222284A1 (en) * 2018-05-14 2019-11-21 The Broad Institute, Inc. In situ cell screening methods and systems
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
CN112673099A (zh) 2018-07-12 2021-04-16 特温斯特兰德生物科学有限公司 用于表征基因组编辑、克隆扩增的方法和试剂及相关应用
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
EP3830289B1 (en) 2018-08-03 2026-02-11 10X Genomics, Inc. Methods to minimize barcode exchange
WO2020041148A1 (en) 2018-08-20 2020-02-27 10X Genomics, Inc. Methods and systems for detection of protein-dna interactions using proximity ligation
US12065688B2 (en) 2018-08-20 2024-08-20 10X Genomics, Inc. Compositions and methods for cellular processing
US11639517B2 (en) 2018-10-01 2023-05-02 Becton, Dickinson And Company Determining 5′ transcript sequences
WO2020081543A1 (en) * 2018-10-16 2020-04-23 The Broad Institute, Inc. Methods of scaling computational genomics with specialized architectures for highly parallelized computations and uses thereof
CN113195735B (zh) * 2018-10-19 2025-03-04 阿科亚生物科学股份有限公司 共存的受体编码核酸区段的检测
EP3877520B1 (en) 2018-11-08 2025-03-19 Becton, Dickson And Company Whole transcriptome analysis of single cells using random priming
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
US11492660B2 (en) 2018-12-13 2022-11-08 Becton, Dickinson And Company Selective extension in single cell whole transcriptome 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
WO2020150356A1 (en) 2019-01-16 2020-07-23 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
US12275993B2 (en) 2019-02-12 2025-04-15 10X Genomics, Inc. Analysis of nucleic acid sequences
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
WO2020167862A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transfer of reagents between droplets
WO2020167866A1 (en) 2019-02-12 2020-08-20 10X Genomics, Inc. Systems and methods for transposon loading
CN118979095A (zh) 2019-02-12 2024-11-19 10X基因组学有限公司 用于加工核酸分子的方法
US11851683B1 (en) 2019-02-12 2023-12-26 10X Genomics, Inc. Methods and systems for selective analysis of cellular samples
US12071617B2 (en) 2019-02-14 2024-08-27 Becton, Dickinson And Company Hybrid targeted and whole transcriptome amplification
US11655499B1 (en) 2019-02-25 2023-05-23 10X Genomics, Inc. Detection of sequence elements in nucleic acid molecules
WO2020185791A1 (en) 2019-03-11 2020-09-17 10X Genomics, Inc. Systems and methods for processing optically tagged beads
WO2020191365A1 (en) 2019-03-21 2020-09-24 Gigamune, Inc. Engineered cells expressing anti-viral t cell receptors and methods of use thereof
WO2020214642A1 (en) 2019-04-19 2020-10-22 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
WO2021003255A1 (en) 2019-07-01 2021-01-07 Mission Bio Method and apparatus to normalize quantitative readouts in single-cell experiments
EP4707405A2 (en) 2019-07-22 2026-03-11 Becton, Dickinson and Company Single cell chromatin immunoprecipitation sequencing assay
US12235262B1 (en) 2019-09-09 2025-02-25 10X Genomics, Inc. Methods and systems for single cell protein analysis
EP4037815A4 (en) * 2019-10-05 2024-01-24 Mission Bio, Inc. METHODS, SYSTEMS AND APPARATUS ASSOCIATED WITH COPY NUMBER VARIATIONS AND SINGLE NUCLEOTIDE VARIATIONS DETECTED SIMULTANEOUSLY IN SINGLE CELLS
EP4055160B1 (en) 2019-11-08 2024-04-10 Becton Dickinson and Company Using random priming to obtain full-length v(d)j information for immune repertoire sequencing
WO2021133842A1 (en) 2019-12-23 2021-07-01 10X Genomics, Inc. Compositions and methods for using fixed biological samples in partition-based assays
CN115135984B (zh) 2019-12-23 2025-12-23 10X基因组学有限公司 可逆固定试剂及其使用方法
WO2021146207A1 (en) 2020-01-13 2021-07-22 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
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
EP4097228B1 (en) 2020-01-29 2024-08-14 Becton, Dickinson and Company Barcoded wells for spatial mapping of single cells through sequencing
US11835462B2 (en) 2020-02-11 2023-12-05 10X Genomics, Inc. Methods and compositions for partitioning a biological sample
US12449419B1 (en) 2020-02-12 2025-10-21 10X Genomics, Inc. Methods for detecting binding of peptide-MHC monomers to T cells
WO2021163630A1 (en) 2020-02-13 2021-08-19 10X Genomics, Inc. Systems and methods for joint interactive visualization of gene expression and dna chromatin accessibility
US20230084773A1 (en) * 2020-02-25 2023-03-16 The Regents Of The University Of California Methods of detecting somatic mutations
WO2021173719A1 (en) 2020-02-25 2021-09-02 Becton, Dickinson And Company Bi-specific probes to enable the use of single-cell samples as single color compensation control
US11926863B1 (en) 2020-02-27 2024-03-12 10X Genomics, Inc. Solid state single cell method for analyzing fixed biological cells
EP4121016A4 (en) 2020-03-16 2024-09-11 Fluent Biosciences Inc. MULTI-OMICS ANALYSIS IN MONODISPERSED DROPLETS
WO2021195169A1 (en) 2020-03-24 2021-09-30 Fluent Biosciences Inc. Viral detection using template emulsification
US11851700B1 (en) 2020-05-13 2023-12-26 10X Genomics, Inc. Methods, kits, and compositions for processing extracellular molecules
WO2021231779A1 (en) 2020-05-14 2021-11-18 Becton, Dickinson And Company Primers for immune repertoire profiling
ES2989052T3 (es) 2020-05-22 2024-11-25 10X Genomics Inc Medición espacio-temporal simultánea de la expresión génica y la actividad celular
JP7423101B2 (ja) * 2020-05-29 2024-01-29 国立研究開発法人理化学研究所 細胞集団の処理方法および細胞集団に含まれる遺伝子の分析方法
CN116249785A (zh) 2020-06-02 2023-06-09 10X基因组学有限公司 用于抗原-受体的空间转录组学
US12265079B1 (en) 2020-06-02 2025-04-01 10X Genomics, Inc. Systems and methods for detecting analytes from captured single biological particles
ES2987035T3 (es) 2020-06-02 2024-11-13 Becton Dickinson Co Oligonucleótidos y perlas para el ensayo de expresión génica principal 5
AU2021283174A1 (en) 2020-06-02 2023-01-05 10X Genomics, Inc. Nucleic acid library methods
US11932901B2 (en) 2020-07-13 2024-03-19 Becton, Dickinson And Company Target enrichment using nucleic acid probes for scRNAseq
JP2023534028A (ja) 2020-07-15 2023-08-07 フルーエント バイオサイエンシーズ インコーポレイテッド 段階的ライゲーション用オリゴ
WO2022026909A1 (en) 2020-07-31 2022-02-03 Becton, Dickinson And Company Single cell assay for transposase-accessible chromatin
US11492662B2 (en) 2020-08-06 2022-11-08 Singular Genomics Systems, Inc. Methods for in situ transcriptomics and proteomics
US11434525B2 (en) 2020-08-06 2022-09-06 Singular Genomics Systems, Inc. Spatial sequencing
US12084715B1 (en) 2020-11-05 2024-09-10 10X Genomics, Inc. Methods and systems for reducing artifactual antisense products
US12480158B1 (en) 2020-11-05 2025-11-25 10X Genomics, Inc. Methods and systems for processing polynucleotides
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
US12398262B1 (en) 2021-01-22 2025-08-26 10X Genomics, Inc. Triblock copolymer-based cell stabilization and fixation system and methods of use thereof
EP4298236A4 (en) * 2021-02-23 2025-02-26 The Broad Institute, Inc. High-throughput assessment of exogenous polynucleotide- or polypeptide-mediated transcriptome perturbations
EP4298244A1 (en) 2021-02-23 2024-01-03 10X Genomics, Inc. Probe-based analysis of nucleic acids and proteins
WO2022235764A1 (en) 2021-05-05 2022-11-10 Singular Genomics Systems, Inc. Multiomic analysis device and methods of use thereof
US12485420B2 (en) 2021-06-03 2025-12-02 Verily Life Sciences Llc Systems and methods for multi-junction particle sorting in injection-molded articles
WO2023287980A1 (en) * 2021-07-15 2023-01-19 Fluent Biosciences Inc. Decentralized workflows for single cell analysis
EP4448787A1 (en) * 2021-12-13 2024-10-23 Cornell University Genotyping of targeted loci with single-cell chromatin accessibility
US12534721B2 (en) 2022-10-06 2026-01-27 Illumina, Inc. Quantitative detection and analysis of moleculesip
AU2024210333A1 (en) 2023-01-20 2025-07-17 Gigagen, Inc. High-throughput analysis of antibody binding and specificity
WO2024249961A1 (en) 2023-06-01 2024-12-05 Singular Genomics Systems, Inc. Methods and probes for detecting polynucleotide sequences in cells and tissues
CN118932067B (zh) * 2024-09-12 2025-10-03 上海信诺佰世医学检验有限公司 Igh基因克隆型检测多重pcr引物组、设计方法及应用

Family Cites Families (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH06509473A (ja) 1991-08-10 1994-10-27 メディカル・リサーチ・カウンシル 細胞個体群の処理
US6013445A (en) * 1996-06-06 2000-01-11 Lynx Therapeutics, Inc. Massively parallel signature sequencing by ligation of encoded adaptors
DE60131903T2 (de) 2000-10-24 2008-11-27 The Board of Trustees of the Leland S. Stanford Junior University, Palo Alto Direkte multiplex charakterisierung von genomischer dna
GB0128153D0 (en) 2001-11-23 2002-01-16 Bayer Ag Profiling of the immune gene repertoire
TWI333977B (en) * 2003-09-18 2010-12-01 Symphogen As Method for linking sequences of interest
US20050221357A1 (en) 2003-09-19 2005-10-06 Mark Shannon Normalization of gene expression data
AU2005325801A1 (en) 2005-01-31 2006-08-03 Ablynx N.V. Method for generating variable domain sequences of heavy chain antibodies
US20060178835A1 (en) 2005-02-10 2006-08-10 Applera Corporation Normalization methods for genotyping analysis
US7497588B2 (en) * 2005-07-14 2009-03-03 International Development Corporation Store display for solar lamp
US8268564B2 (en) * 2007-09-26 2012-09-18 President And Fellows Of Harvard College Methods and applications for stitched DNA barcodes
WO2009049889A1 (en) * 2007-10-16 2009-04-23 Roche Diagnostics Gmbh High resolution, high throughput hla genotyping by clonal sequencing
WO2010053587A2 (en) 2008-11-07 2010-05-14 Mlc Dx Incorporated Methods of monitoring conditions by sequence analysis
JP2012525147A (ja) 2009-04-30 2012-10-22 グッド スタート ジェネティクス， インコーポレイテッド 遺伝マーカーを評価するための方法および組成物
ES2740802T3 (es) * 2010-05-06 2020-02-06 Adaptive Biotechnologies Corp Monitorización del estadio de neoplasia linfoide usando perfiles de clonotipo
CA2799746C (en) 2010-05-17 2020-11-24 Sai Reddy Rapid isolation of monoclonal antibodies from animals
EP3561073A1 (en) 2010-10-08 2019-10-30 President and Fellows of Harvard College High-throughput immune sequencing
ES2731460T3 (es) * 2010-10-08 2019-11-15 Harvard College Alto rendimiento de células individuales con código de barra
DK2652155T3 (da) * 2010-12-16 2017-02-13 Gigagen Inc Fremgangsmåder til massiv parallelanalyse af nukleinsyrer i enkeltceller
WO2012104851A1 (en) * 2011-01-31 2012-08-09 Yeda Research And Development Co. Ltd. Methods of diagnosing disease using overlap extension pcr
WO2013109935A1 (en) 2012-01-20 2013-07-25 Gigagen, Inc. Method for analysis of immune variable sequences
WO2013112655A1 (en) 2012-01-24 2013-08-01 Gigagen, Inc. Method for correction of bias in multiplexed amplification
WO2013188772A1 (en) 2012-06-14 2013-12-19 Gigagen, Inc. Methods for antibody and cell therapeutic discovery
WO2014004124A2 (en) 2012-06-14 2014-01-03 Gigagen, Inc. Method for amplification gene fusion complexes
ES2774165T3 (es) 2012-06-15 2020-07-17 Univ Texas Secuenciación de alto rendimiento de múltiples transcritos de una única célula
CA2900543C (en) 2013-02-08 2023-01-31 10X Genomics, Inc. Partitioning and processing of analytes and other species

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