Microfluidic-free encapsulation of bacteria for isothermal DNA amplification

Autores
Fonseca, A.; Bilen, Marcos Fabian; Olivares, María Laura; Marengo, Robinson Cristian; Berli, Claudio Luis Alberto; Borio, Cristina Silvia
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Linking genotype (a nucleic acid that can be replicated) and phenotype (a functional trait, such as a binding or catalytic or regulatory activity) in a unique compartment is a key aspect of protein functional analysis, because emulates the natural genotype-to-phenotype linkage that results from the compartmentalization of genes in cells.This connection is easy to obtain when the number of samples is below 100, but when the number of samples rises to the order of millions, a technical problem appears. A solution to this limitation is the miniaturization and parallelization of the process, associated with the formation of aqueous nanocompartments, like aqueous droplets in oil phases. This methodology converts the analysis into ultra-high-throughput, since is possible to obtain compartments of small sizes as 100 mm and volumes of nearly 1 nanolitre. This high capacity (>1010 droplets in 1 ml of emulsion), the ease of preparing emulsions and their high stability over a broad range of temperatures, pH and salt concentrations, makes this methodology ideal for compartmentalizing biochemical and genetic assays.In this sense, the nanocompartmentalization of bacteria is an optimal tool for in vitro evolution analysis, with the only condition that it is possible to confine a single bacteria per compartment. To do this in a conventional system the water-in-oil emulsion formation parameters, must be considered.In this study, parameters like bacterial DO600 and vortex speed where evaluated in order to obtain particles of average size of 100mm containing a bacteria or none inside. Bacterias expressing a DNA polymerase were used to study the isothermal DNA amplification inside these particles, by DNA recovery and fluorescence detection. Here we also report on preliminary studies towards microfluidic generation of microdroplets. A PMMA chip with a cross-junction was used to obtain W/O emulsion, with fluids fed by syringe pumps.
Fil: Fonseca, A.. Universidad Nacional de Quilmes; Argentina
Fil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Olivares, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Marengo, Robinson Cristian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Berli, Claudio Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Borio, Cristina Silvia. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
II Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica Argentina
Córodoba
Argentina
Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía, Física y Computación
Materia
ENCAPSULATION
DNA AMPLIFICATION
MICROFLUIDIC
MICROPARTICLES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/136221
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/136221
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Microfluidic-free encapsulation of bacteria for isothermal DNA amplificationFonseca, A.Bilen, Marcos FabianOlivares, María LauraMarengo, Robinson CristianBerli, Claudio Luis AlbertoBorio, Cristina SilviaENCAPSULATIONDNA AMPLIFICATIONMICROFLUIDICMICROPARTICLEShttps://purl.org/becyt/ford/2.10https://purl.org/becyt/ford/2Linking genotype (a nucleic acid that can be replicated) and phenotype (a functional trait, such as a binding or catalytic or regulatory activity) in a unique compartment is a key aspect of protein functional analysis, because emulates the natural genotype-to-phenotype linkage that results from the compartmentalization of genes in cells.This connection is easy to obtain when the number of samples is below 100, but when the number of samples rises to the order of millions, a technical problem appears. A solution to this limitation is the miniaturization and parallelization of the process, associated with the formation of aqueous nanocompartments, like aqueous droplets in oil phases. This methodology converts the analysis into ultra-high-throughput, since is possible to obtain compartments of small sizes as 100 mm and volumes of nearly 1 nanolitre. This high capacity (>1010 droplets in 1 ml of emulsion), the ease of preparing emulsions and their high stability over a broad range of temperatures, pH and salt concentrations, makes this methodology ideal for compartmentalizing biochemical and genetic assays.In this sense, the nanocompartmentalization of bacteria is an optimal tool for in vitro evolution analysis, with the only condition that it is possible to confine a single bacteria per compartment. To do this in a conventional system the water-in-oil emulsion formation parameters, must be considered.In this study, parameters like bacterial DO600 and vortex speed where evaluated in order to obtain particles of average size of 100mm containing a bacteria or none inside. Bacterias expressing a DNA polymerase were used to study the isothermal DNA amplification inside these particles, by DNA recovery and fluorescence detection. Here we also report on preliminary studies towards microfluidic generation of microdroplets. A PMMA chip with a cross-junction was used to obtain W/O emulsion, with fluids fed by syringe pumps.Fil: Fonseca, A.. Universidad Nacional de Quilmes; ArgentinaFil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Olivares, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Marengo, Robinson Cristian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Berli, Claudio Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Borio, Cristina Silvia. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaII Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica ArgentinaCórodobaArgentinaUniversidad Nacional de Córdoba. Facultad de Matemática, Astronomía, Física y ComputaciónUniversidad Nacional de CórdobaMarconi, Verónica I.Banchio, Adolfo J.2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/136221Microfluidic-free encapsulation of bacteria for isothermal DNA amplification; II Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica Argentina; Córodoba; Argentina; 2019; 1-6978-987-779-009-2CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mfarg.org/book-of-abstractsInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:20:16Zoai:ri.conicet.gov.ar:11336/136221instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:20:17.308CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
title Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
spellingShingle Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
Fonseca, A.
ENCAPSULATION
DNA AMPLIFICATION
MICROFLUIDIC
MICROPARTICLES
title_short Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
title_full Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
title_fullStr Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
title_full_unstemmed Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
title_sort Microfluidic-free encapsulation of bacteria for isothermal DNA amplification
dc.creator.none.fl_str_mv Fonseca, A.
Bilen, Marcos Fabian
Olivares, María Laura
Marengo, Robinson Cristian
Berli, Claudio Luis Alberto
Borio, Cristina Silvia
author Fonseca, A.
author_facet Fonseca, A.
Bilen, Marcos Fabian
Olivares, María Laura
Marengo, Robinson Cristian
Berli, Claudio Luis Alberto
Borio, Cristina Silvia
author_role author
author2 Bilen, Marcos Fabian
Olivares, María Laura
Marengo, Robinson Cristian
Berli, Claudio Luis Alberto
Borio, Cristina Silvia
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Marconi, Verónica I.
Banchio, Adolfo J.
dc.subject.none.fl_str_mv ENCAPSULATION
DNA AMPLIFICATION
MICROFLUIDIC
MICROPARTICLES
topic ENCAPSULATION
DNA AMPLIFICATION
MICROFLUIDIC
MICROPARTICLES
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.10
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Linking genotype (a nucleic acid that can be replicated) and phenotype (a functional trait, such as a binding or catalytic or regulatory activity) in a unique compartment is a key aspect of protein functional analysis, because emulates the natural genotype-to-phenotype linkage that results from the compartmentalization of genes in cells.This connection is easy to obtain when the number of samples is below 100, but when the number of samples rises to the order of millions, a technical problem appears. A solution to this limitation is the miniaturization and parallelization of the process, associated with the formation of aqueous nanocompartments, like aqueous droplets in oil phases. This methodology converts the analysis into ultra-high-throughput, since is possible to obtain compartments of small sizes as 100 mm and volumes of nearly 1 nanolitre. This high capacity (>1010 droplets in 1 ml of emulsion), the ease of preparing emulsions and their high stability over a broad range of temperatures, pH and salt concentrations, makes this methodology ideal for compartmentalizing biochemical and genetic assays.In this sense, the nanocompartmentalization of bacteria is an optimal tool for in vitro evolution analysis, with the only condition that it is possible to confine a single bacteria per compartment. To do this in a conventional system the water-in-oil emulsion formation parameters, must be considered.In this study, parameters like bacterial DO600 and vortex speed where evaluated in order to obtain particles of average size of 100mm containing a bacteria or none inside. Bacterias expressing a DNA polymerase were used to study the isothermal DNA amplification inside these particles, by DNA recovery and fluorescence detection. Here we also report on preliminary studies towards microfluidic generation of microdroplets. A PMMA chip with a cross-junction was used to obtain W/O emulsion, with fluids fed by syringe pumps.
Fil: Fonseca, A.. Universidad Nacional de Quilmes; Argentina
Fil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Olivares, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Marengo, Robinson Cristian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Berli, Claudio Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentina
Fil: Borio, Cristina Silvia. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
II Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica Argentina
Córodoba
Argentina
Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía, Física y Computación
description Linking genotype (a nucleic acid that can be replicated) and phenotype (a functional trait, such as a binding or catalytic or regulatory activity) in a unique compartment is a key aspect of protein functional analysis, because emulates the natural genotype-to-phenotype linkage that results from the compartmentalization of genes in cells.This connection is easy to obtain when the number of samples is below 100, but when the number of samples rises to the order of millions, a technical problem appears. A solution to this limitation is the miniaturization and parallelization of the process, associated with the formation of aqueous nanocompartments, like aqueous droplets in oil phases. This methodology converts the analysis into ultra-high-throughput, since is possible to obtain compartments of small sizes as 100 mm and volumes of nearly 1 nanolitre. This high capacity (>1010 droplets in 1 ml of emulsion), the ease of preparing emulsions and their high stability over a broad range of temperatures, pH and salt concentrations, makes this methodology ideal for compartmentalizing biochemical and genetic assays.In this sense, the nanocompartmentalization of bacteria is an optimal tool for in vitro evolution analysis, with the only condition that it is possible to confine a single bacteria per compartment. To do this in a conventional system the water-in-oil emulsion formation parameters, must be considered.In this study, parameters like bacterial DO600 and vortex speed where evaluated in order to obtain particles of average size of 100mm containing a bacteria or none inside. Bacterias expressing a DNA polymerase were used to study the isothermal DNA amplification inside these particles, by DNA recovery and fluorescence detection. Here we also report on preliminary studies towards microfluidic generation of microdroplets. A PMMA chip with a cross-junction was used to obtain W/O emulsion, with fluids fed by syringe pumps.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/136221
Microfluidic-free encapsulation of bacteria for isothermal DNA amplification; II Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica Argentina; Córodoba; Argentina; 2019; 1-6
978-987-779-009-2
CONICET Digital
CONICET
url http://hdl.handle.net/11336/136221
identifier_str_mv Microfluidic-free encapsulation of bacteria for isothermal DNA amplification; II Brazil-Argentine Microfluidics Congress y V Congreso de Microfluídica Argentina; Córodoba; Argentina; 2019; 1-6
978-987-779-009-2
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.mfarg.org/book-of-abstracts
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Universidad Nacional de Córdoba
publisher.none.fl_str_mv Universidad Nacional de Córdoba
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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