Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples

Detalhes bibliográficos
Autor(a) principal: Churro, Catarina
Data de Publicação: 2015
Outros Autores: Valério, Elisabete, Pereira, Paulo, Vasconcelos, Vitor
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.18/3775
Resumo: Publicaciones del III Congreso Ibérico de Cianotoxinas (Blanes, 2013)
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spelling Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samplesCyanobacteriaGlutaraldehydeFormaldehydeLugol’s iodineReal-time qPCRPlanktothrixÁgua e SoloPublicaciones del III Congreso Ibérico de Cianotoxinas (Blanes, 2013)The study and monitoring of cyanobacterial blooms often involves the use of preserved samples to avoid cellular degradation. However, preserved samples may not be suitable for molecular biology studies because preservation methods can interfere with DNA quality/quantity. Real-time quantitative PCR analysis (qPCR) has been widely applied in molecular analysis and is considered a promising method for monitoring purposes. This study intended to evaluate the applicability of the real-time qPCR technique in samples that were subjected to different methods of preservation: (1) 15% Lugol’s iodine solution (2) 4% formaldehyde and (3) 25% glutaraldehyde. The ability to amplify and quantify DNA extracted from Planktothrix agardhii was assessed using the rpoC1 gene as the target fragment in both raw water samples and in vitro cultures. No reliable DNA amplification was obtained from glutaraldehyde-preserved samples. Successful amplification was obtained from Lugol’s and formaldehyde-preserved samples. In this case, however, the quantification that was achieved by real-time PCR cannot be used to infer cell numbers, because the Ct values that were obtained from the Lugol’s and formaldehydepreserved samples were significantly higher than the Ct values that were obtained from the unpreserved samples. Therefore real-time PCR can be used for the detection and identification of cyanobacteria in preserved samples but no reliable cell quantification can be performed using this method.This research was supported by the Fundação para Ciência e Tecnologia, Portugal (FCT) through the project PPCDT/AMB/67075/2006. The authors also acknowledge the PhD research grant SFRH/BD65706/2009 to C. Churro and the Post-Doc research grant SFRH/BPD/75922/2011 to E. Valério.Asociación Ibérica de LimnologíaRepositório Científico do Instituto Nacional de SaúdeChurro, CatarinaValério, ElisabetePereira, PauloVasconcelos, Vitor2016-04-06T11:43:19Z20152015-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/3775engLimnetica. 2015;34(1):173-860213-8409info: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:RCAAP2023-07-20T15:39:58Zoai:repositorio.insa.pt:10400.18/3775Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:38:35.215195Repositório 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 Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
title Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
spellingShingle Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
Churro, Catarina
Cyanobacteria
Glutaraldehyde
Formaldehyde
Lugol’s iodine
Real-time qPCR
Planktothrix
Água e Solo
title_short Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
title_full Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
title_fullStr Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
title_full_unstemmed Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
title_sort Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples
author Churro, Catarina
author_facet Churro, Catarina
Valério, Elisabete
Pereira, Paulo
Vasconcelos, Vitor
author_role author
author2 Valério, Elisabete
Pereira, Paulo
Vasconcelos, Vitor
author2_role author
author
author
dc.contributor.none.fl_str_mv Repositório Científico do Instituto Nacional de Saúde
dc.contributor.author.fl_str_mv Churro, Catarina
Valério, Elisabete
Pereira, Paulo
Vasconcelos, Vitor
dc.subject.por.fl_str_mv Cyanobacteria
Glutaraldehyde
Formaldehyde
Lugol’s iodine
Real-time qPCR
Planktothrix
Água e Solo
topic Cyanobacteria
Glutaraldehyde
Formaldehyde
Lugol’s iodine
Real-time qPCR
Planktothrix
Água e Solo
description Publicaciones del III Congreso Ibérico de Cianotoxinas (Blanes, 2013)
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-01-01T00:00:00Z
2016-04-06T11:43:19Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.18/3775
url http://hdl.handle.net/10400.18/3775
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Limnetica. 2015;34(1):173-86
0213-8409
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 Asociación Ibérica de Limnología
publisher.none.fl_str_mv Asociación Ibérica de Limnología
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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