Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/3774 |
Resumo: | Molecular studies on cyanobacteria often involve filtering and freezing of samples leading to loss of cell morphological features. Methanol is often used in preservation of biological materials in association with other fixatives. This study evaluates the application of methanol in the preservation of DNA for molecular studies as well as for the preservation of cell morphology for morphometric analysis in filamentous cyanobacteria. In the present study, both culture and environmental bloom samples were preserved using a cold methanol dehydration series (50, 70, and 100 %) and stored at −20 °C for up to 2 years. The DNA quantity and quality, nucleotide sequence retrieval, and real-time PCR quantification were analyzed over time. Morphometric cell analysis was performed on preserved samples. Results show that the DNA extracted from samples preserved up to 6 months was successfully quantified by real-time PCR. After that period, the DNA quantity decreased with the preservation time. Nevertheless, we were able to detect/amplify the target fragment in samples preserved up to 2 years. The DNA sequence and cell morphology were also maintained during the preservation time. Thus, methanol preservation is an adequate method to preserve molecular information and morphological features after long storage periods. |
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Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhiiCyanobacteriaLugol’s iodineMethanolPreservationReal-time qPCRrpoC1Água e SoloMolecular studies on cyanobacteria often involve filtering and freezing of samples leading to loss of cell morphological features. Methanol is often used in preservation of biological materials in association with other fixatives. This study evaluates the application of methanol in the preservation of DNA for molecular studies as well as for the preservation of cell morphology for morphometric analysis in filamentous cyanobacteria. In the present study, both culture and environmental bloom samples were preserved using a cold methanol dehydration series (50, 70, and 100 %) and stored at −20 °C for up to 2 years. The DNA quantity and quality, nucleotide sequence retrieval, and real-time PCR quantification were analyzed over time. Morphometric cell analysis was performed on preserved samples. Results show that the DNA extracted from samples preserved up to 6 months was successfully quantified by real-time PCR. After that period, the DNA quantity decreased with the preservation time. Nevertheless, we were able to detect/amplify the target fragment in samples preserved up to 2 years. The DNA sequence and cell morphology were also maintained during the preservation time. Thus, methanol preservation is an adequate method to preserve molecular information and morphological features after long storage periods.This research was partially supported by the European Regional Development Fund (ERDF) through the COMPETE - Operational Competitiveness Program - and FCT - Foundation for Science and Technology national funds through projects PPCDT/AMB/67075/2006 and PEst-C/MAR/LA0015/2013, the Post-Doc research grant SFRH/BPD/75922/2011 to E. Valério and Ph.D. research grant SFRH/BD65706/2009 to C. Churro.Springer VerlagRepositório Científico do Instituto Nacional de SaúdeChurro, CatarinaValério, ElisabeteVieira, LuísPereira, PauloVasconcelos, Vitor2016-11-01T01:30:09Z2015-10-292015-10-29T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/3774engJ. Appl. Phycol. 2015. Epub 2015 Oct 29. doi: 10.1007/s10811-015-0732-0).0921-897110.1007/s10811-015-0732-0info:eu-repo/semantics/embargoedAccessreponame: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:10400.18/3774Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:38:35.270045Repositório 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 |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
title |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
spellingShingle |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii Churro, Catarina Cyanobacteria Lugol’s iodine Methanol Preservation Real-time qPCR rpoC1 Água e Solo |
title_short |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
title_full |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
title_fullStr |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
title_full_unstemmed |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
title_sort |
Evaluation of methanol preservation for molecular and morphological studies in cyanobacteria using Planktothrix agardhii |
author |
Churro, Catarina |
author_facet |
Churro, Catarina Valério, Elisabete Vieira, Luís Pereira, Paulo Vasconcelos, Vitor |
author_role |
author |
author2 |
Valério, Elisabete Vieira, Luís Pereira, Paulo Vasconcelos, Vitor |
author2_role |
author 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 Vieira, Luís Pereira, Paulo Vasconcelos, Vitor |
dc.subject.por.fl_str_mv |
Cyanobacteria Lugol’s iodine Methanol Preservation Real-time qPCR rpoC1 Água e Solo |
topic |
Cyanobacteria Lugol’s iodine Methanol Preservation Real-time qPCR rpoC1 Água e Solo |
description |
Molecular studies on cyanobacteria often involve filtering and freezing of samples leading to loss of cell morphological features. Methanol is often used in preservation of biological materials in association with other fixatives. This study evaluates the application of methanol in the preservation of DNA for molecular studies as well as for the preservation of cell morphology for morphometric analysis in filamentous cyanobacteria. In the present study, both culture and environmental bloom samples were preserved using a cold methanol dehydration series (50, 70, and 100 %) and stored at −20 °C for up to 2 years. The DNA quantity and quality, nucleotide sequence retrieval, and real-time PCR quantification were analyzed over time. Morphometric cell analysis was performed on preserved samples. Results show that the DNA extracted from samples preserved up to 6 months was successfully quantified by real-time PCR. After that period, the DNA quantity decreased with the preservation time. Nevertheless, we were able to detect/amplify the target fragment in samples preserved up to 2 years. The DNA sequence and cell morphology were also maintained during the preservation time. Thus, methanol preservation is an adequate method to preserve molecular information and morphological features after long storage periods. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-10-29 2015-10-29T00:00:00Z 2016-11-01T01:30:09Z |
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/10400.18/3774 |
url |
http://hdl.handle.net/10400.18/3774 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
J. Appl. Phycol. 2015. Epub 2015 Oct 29. doi: 10.1007/s10811-015-0732-0). 0921-8971 10.1007/s10811-015-0732-0 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Springer Verlag |
publisher.none.fl_str_mv |
Springer Verlag |
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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1799132123632762880 |