A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/99431 |
Resumo: | Digital Microfluidics (DMF) is an innovative technology for liquid manipulation at microliter- to picoliter-scale, with tremendous potential of application in biosensing. DMF allows maneuvering single droplets over an electrode array, by means of electrowetting-on-dielectric (EWOD), that allows changing the contact angle of a droplet over a dielectric. Each droplet is thus considered a microreactor, with an unparalleled potential to perform chemical and biological reactions. Several aspects inherent to DMF platforms, such as multiplex assay capability and integration capability, make them promising for lab-on-chip and point-of-care (PoC) applications, e.g. DNA amplification assays or disease detection. DNA detection strategies for PoC have been profiting from recent development of isothermal amplification schemes, of which Loop-mediated Isothermal Amplification (LAMP) is a major methodology, allowing a 109-fold amplification efficiency in one hour. Here, I demonstrate for the first time the effective coupling of DMF and LAMP, resulting in a DMF device capable of performing LAMP reactions. This novel DMF platform has been developed and characterised, which allows successful amplification of a c-Myc gene fragment by LAMP. Precise temperature control is achieved by using a transparent heating element, connected to a looping feedback control system. This platform is able to amplify just 0.5 ng/μL of the target DNA, in only 45 minutes, for a device temperature of 65 °C and a reaction volume of 1.62 μL, one of the lowest volumes ever reported. Moreover, the electrophoretic analysis indicates that the amplification efficiency of the on-chip LAMP is considerably higher than that from the bench-top reaction. |
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A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNADigital MicrofluidicsLoop-mediated Isothermal Amplificationc-Myclab-on-chippoint-of-care diagnosticsDomínio/Área Científica::Engenharia e Tecnologia::NanotecnologiaDigital Microfluidics (DMF) is an innovative technology for liquid manipulation at microliter- to picoliter-scale, with tremendous potential of application in biosensing. DMF allows maneuvering single droplets over an electrode array, by means of electrowetting-on-dielectric (EWOD), that allows changing the contact angle of a droplet over a dielectric. Each droplet is thus considered a microreactor, with an unparalleled potential to perform chemical and biological reactions. Several aspects inherent to DMF platforms, such as multiplex assay capability and integration capability, make them promising for lab-on-chip and point-of-care (PoC) applications, e.g. DNA amplification assays or disease detection. DNA detection strategies for PoC have been profiting from recent development of isothermal amplification schemes, of which Loop-mediated Isothermal Amplification (LAMP) is a major methodology, allowing a 109-fold amplification efficiency in one hour. Here, I demonstrate for the first time the effective coupling of DMF and LAMP, resulting in a DMF device capable of performing LAMP reactions. This novel DMF platform has been developed and characterised, which allows successful amplification of a c-Myc gene fragment by LAMP. Precise temperature control is achieved by using a transparent heating element, connected to a looping feedback control system. This platform is able to amplify just 0.5 ng/μL of the target DNA, in only 45 minutes, for a device temperature of 65 °C and a reaction volume of 1.62 μL, one of the lowest volumes ever reported. Moreover, the electrophoretic analysis indicates that the amplification efficiency of the on-chip LAMP is considerably higher than that from the bench-top reaction.Igreja, RuiBaptista, PedroRUNCoelho, Beatriz Jorge2020-06-16T14:51:45Z2016-1220162016-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/99431enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-05-22T17:46:02Zoai:run.unl.pt:10362/99431Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-05-22T17:46:02Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
title |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
spellingShingle |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA Coelho, Beatriz Jorge Digital Microfluidics Loop-mediated Isothermal Amplification c-Myc lab-on-chip point-of-care diagnostics Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
title_short |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
title_full |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
title_fullStr |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
title_full_unstemmed |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
title_sort |
A Digital Microfluidics Platform for Loop-Mediated Isothermal Amplification of DNA |
author |
Coelho, Beatriz Jorge |
author_facet |
Coelho, Beatriz Jorge |
author_role |
author |
dc.contributor.none.fl_str_mv |
Igreja, Rui Baptista, Pedro RUN |
dc.contributor.author.fl_str_mv |
Coelho, Beatriz Jorge |
dc.subject.por.fl_str_mv |
Digital Microfluidics Loop-mediated Isothermal Amplification c-Myc lab-on-chip point-of-care diagnostics Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
topic |
Digital Microfluidics Loop-mediated Isothermal Amplification c-Myc lab-on-chip point-of-care diagnostics Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
description |
Digital Microfluidics (DMF) is an innovative technology for liquid manipulation at microliter- to picoliter-scale, with tremendous potential of application in biosensing. DMF allows maneuvering single droplets over an electrode array, by means of electrowetting-on-dielectric (EWOD), that allows changing the contact angle of a droplet over a dielectric. Each droplet is thus considered a microreactor, with an unparalleled potential to perform chemical and biological reactions. Several aspects inherent to DMF platforms, such as multiplex assay capability and integration capability, make them promising for lab-on-chip and point-of-care (PoC) applications, e.g. DNA amplification assays or disease detection. DNA detection strategies for PoC have been profiting from recent development of isothermal amplification schemes, of which Loop-mediated Isothermal Amplification (LAMP) is a major methodology, allowing a 109-fold amplification efficiency in one hour. Here, I demonstrate for the first time the effective coupling of DMF and LAMP, resulting in a DMF device capable of performing LAMP reactions. This novel DMF platform has been developed and characterised, which allows successful amplification of a c-Myc gene fragment by LAMP. Precise temperature control is achieved by using a transparent heating element, connected to a looping feedback control system. This platform is able to amplify just 0.5 ng/μL of the target DNA, in only 45 minutes, for a device temperature of 65 °C and a reaction volume of 1.62 μL, one of the lowest volumes ever reported. Moreover, the electrophoretic analysis indicates that the amplification efficiency of the on-chip LAMP is considerably higher than that from the bench-top reaction. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-12 2016 2016-12-01T00:00:00Z 2020-06-16T14:51:45Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/99431 |
url |
http://hdl.handle.net/10362/99431 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
mluisa.alvim@gmail.com |
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1817545746796249088 |