Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis

Detalhes bibliográficos
Autor(a) principal: Scatena, Mário Pinzan [UNESP]
Data de Publicação: 2008
Outros Autores: Morielle-Versute, Eliana [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/S1415-47572008000100027
http://hdl.handle.net/11449/22519
Resumo: To establish a technique which minimized the effects of fixation on the extraction of DNA from formalin-fixed tissues preserved in scientific collections we extracted DNA samples from fixed tissues using different methods and evaluated the effect of the different procedures on PCR and sequencing analysis. We investigated muscle and liver tissues from museum specimens of five species of fruit-eating (frugivorous) bats of the Neotropical genus Artibeus (Chiroptera, Phyllostomidae): A. fimbriatus, A. lituratus, A. jamaicensis, A. obscurus, and A. planirostris. The results indicated that treatment of tissues in buffered solutions at neutral pH and about 37 °C for at least four days improves the quality and quantity of extracted DNA and the quality of the amplification and sequencing products. However, the comparison between the performance of DNA obtained from fixed and fresh tissues showed that, in spite of the fact that both types of tissue generate reliable sequences for use in phylogenetic analyses, DNA samples from fixed tissues presented a larger rate of errors in the different stages of the study. These results suggest that DNA extracted from formalin-fixed tissue can be used in molecular studies of Neotropical Artibeus bats and that our methodology may be applicable to other animal groups.
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spelling Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysisarchival specimensChiropteraDNA extractionformalin-fixedTo establish a technique which minimized the effects of fixation on the extraction of DNA from formalin-fixed tissues preserved in scientific collections we extracted DNA samples from fixed tissues using different methods and evaluated the effect of the different procedures on PCR and sequencing analysis. We investigated muscle and liver tissues from museum specimens of five species of fruit-eating (frugivorous) bats of the Neotropical genus Artibeus (Chiroptera, Phyllostomidae): A. fimbriatus, A. lituratus, A. jamaicensis, A. obscurus, and A. planirostris. The results indicated that treatment of tissues in buffered solutions at neutral pH and about 37 °C for at least four days improves the quality and quantity of extracted DNA and the quality of the amplification and sequencing products. However, the comparison between the performance of DNA obtained from fixed and fresh tissues showed that, in spite of the fact that both types of tissue generate reliable sequences for use in phylogenetic analyses, DNA samples from fixed tissues presented a larger rate of errors in the different stages of the study. These results suggest that DNA extracted from formalin-fixed tissue can be used in molecular studies of Neotropical Artibeus bats and that our methodology may be applicable to other animal groups.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação para o Desenvolvimento da UNESP (FUNDUNESP)Universidade Estadual Paulista Instituto de Biociências, Letras e Ciências Exatas Departamento de Zoologia e BotânicaUniversidade Estadual Paulista Instituto de Biociências, Letras e Ciências Exatas Departamento de Zoologia e BotânicaSociedade Brasileira de GenéticaUniversidade Estadual Paulista (Unesp)Scatena, Mário Pinzan [UNESP]Morielle-Versute, Eliana [UNESP]2014-05-20T14:04:06Z2014-05-20T14:04:06Z2008-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article160-165application/pdfhttp://dx.doi.org/10.1590/S1415-47572008000100027Genetics and Molecular Biology. Sociedade Brasileira de Genética, v. 31, n. 1, p. 160-165, 2008.1415-4757http://hdl.handle.net/11449/2251910.1590/S1415-47572008000100027S1415-47572008000100027WOS:000254487000027S1415-47572008000100027.pdfSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengGenetics and Molecular Biology1.4930,638info:eu-repo/semantics/openAccess2023-10-25T06:08:55Zoai:repositorio.unesp.br:11449/22519Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-25T06:08:55Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
title Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
spellingShingle Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
Scatena, Mário Pinzan [UNESP]
archival specimens
Chiroptera
DNA extraction
formalin-fixed
title_short Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
title_full Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
title_fullStr Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
title_full_unstemmed Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
title_sort Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
author Scatena, Mário Pinzan [UNESP]
author_facet Scatena, Mário Pinzan [UNESP]
Morielle-Versute, Eliana [UNESP]
author_role author
author2 Morielle-Versute, Eliana [UNESP]
author2_role author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Scatena, Mário Pinzan [UNESP]
Morielle-Versute, Eliana [UNESP]
dc.subject.por.fl_str_mv archival specimens
Chiroptera
DNA extraction
formalin-fixed
topic archival specimens
Chiroptera
DNA extraction
formalin-fixed
description To establish a technique which minimized the effects of fixation on the extraction of DNA from formalin-fixed tissues preserved in scientific collections we extracted DNA samples from fixed tissues using different methods and evaluated the effect of the different procedures on PCR and sequencing analysis. We investigated muscle and liver tissues from museum specimens of five species of fruit-eating (frugivorous) bats of the Neotropical genus Artibeus (Chiroptera, Phyllostomidae): A. fimbriatus, A. lituratus, A. jamaicensis, A. obscurus, and A. planirostris. The results indicated that treatment of tissues in buffered solutions at neutral pH and about 37 °C for at least four days improves the quality and quantity of extracted DNA and the quality of the amplification and sequencing products. However, the comparison between the performance of DNA obtained from fixed and fresh tissues showed that, in spite of the fact that both types of tissue generate reliable sequences for use in phylogenetic analyses, DNA samples from fixed tissues presented a larger rate of errors in the different stages of the study. These results suggest that DNA extracted from formalin-fixed tissue can be used in molecular studies of Neotropical Artibeus bats and that our methodology may be applicable to other animal groups.
publishDate 2008
dc.date.none.fl_str_mv 2008-01-01
2014-05-20T14:04:06Z
2014-05-20T14:04:06Z
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://dx.doi.org/10.1590/S1415-47572008000100027
Genetics and Molecular Biology. Sociedade Brasileira de Genética, v. 31, n. 1, p. 160-165, 2008.
1415-4757
http://hdl.handle.net/11449/22519
10.1590/S1415-47572008000100027
S1415-47572008000100027
WOS:000254487000027
S1415-47572008000100027.pdf
url http://dx.doi.org/10.1590/S1415-47572008000100027
http://hdl.handle.net/11449/22519
identifier_str_mv Genetics and Molecular Biology. Sociedade Brasileira de Genética, v. 31, n. 1, p. 160-165, 2008.
1415-4757
10.1590/S1415-47572008000100027
S1415-47572008000100027
WOS:000254487000027
S1415-47572008000100027.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Genetics and Molecular Biology
1.493
0,638
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 160-165
application/pdf
dc.publisher.none.fl_str_mv Sociedade Brasileira de Genética
publisher.none.fl_str_mv Sociedade Brasileira de Genética
dc.source.none.fl_str_mv SciELO
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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