mRNA genotyping by gold nanoprobes

Detalhes bibliográficos
Autor(a) principal: Assunção, Maria Saraiva Mendes
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/6235
Resumo: Dissertação para obtenção do Grau de Mestre em Biotecnologia
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spelling mRNA genotyping by gold nanoprobesBionanotechnologyNon-cross-linking assayGold nanoprobesRNA genotypingBRCA1Dissertação para obtenção do Grau de Mestre em BiotecnologiaBionanotechnology has emerged as a field with great potential for molecular diagnose. Namely, the use of gold nanoparticles has allowed the development of molecular diagnostic methods with greater sensitivity and specificity at a fraction of the cost inherent to conventional techniques. The present work assessed the ability of gold nanoprobes to detect targets with single base differences in RNA molecules following the colorimetric non-cross-linking method. As proof-of-concept, gold nanoprobes were designed, synthesized and characterized to detect three different SNPs (c.2731C>T, c.3232A>G and c.3238G>A) in the BRCA1 gene, a gene associated with inherited breast cancer. Reference materials, susceptible to be used for the calibration of the method, were created by cloning genomic fragments amplified from biological samples containing the sequences of interest in an appropriate vector for subsequent in vitro/in vivo transcription. Initially, the ability of target recognition by the gold nanoprobes was assessed using synthetic oligonucleotides targets alone and spiked-in total RNA of Saccharomyces cerevisiae, and later using transcripts synthetized in vitro/in vivo. This study revealed the capacity of target detection up to 0.25% of complementary target/total RNA, for a final concentration of complementary target of 0.12pmol/μL. It was also possible to detect and discriminate both c.3232A>G SNP alleles using only 0.08pmol/μL of in vitro transcript. For the in vivo transcript samples the results were inconclusive.Faculdade de Ciências e TecnologiaBaptista, PedroDoria, GonçaloRUNAssunção, Maria Saraiva Mendes2011-11-21T15:24:03Z20112011-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/6235enginfo: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:RCAAP2024-03-11T03:37:22Zoai:run.unl.pt:10362/6235Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:16:47.329102Repositório 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 mRNA genotyping by gold nanoprobes
title mRNA genotyping by gold nanoprobes
spellingShingle mRNA genotyping by gold nanoprobes
Assunção, Maria Saraiva Mendes
Bionanotechnology
Non-cross-linking assay
Gold nanoprobes
RNA genotyping
BRCA1
title_short mRNA genotyping by gold nanoprobes
title_full mRNA genotyping by gold nanoprobes
title_fullStr mRNA genotyping by gold nanoprobes
title_full_unstemmed mRNA genotyping by gold nanoprobes
title_sort mRNA genotyping by gold nanoprobes
author Assunção, Maria Saraiva Mendes
author_facet Assunção, Maria Saraiva Mendes
author_role author
dc.contributor.none.fl_str_mv Baptista, Pedro
Doria, Gonçalo
RUN
dc.contributor.author.fl_str_mv Assunção, Maria Saraiva Mendes
dc.subject.por.fl_str_mv Bionanotechnology
Non-cross-linking assay
Gold nanoprobes
RNA genotyping
BRCA1
topic Bionanotechnology
Non-cross-linking assay
Gold nanoprobes
RNA genotyping
BRCA1
description Dissertação para obtenção do Grau de Mestre em Biotecnologia
publishDate 2011
dc.date.none.fl_str_mv 2011-11-21T15:24:03Z
2011
2011-01-01T00:00:00Z
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/10362/6235
url http://hdl.handle.net/10362/6235
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.publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
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
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