Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | |
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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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:29462024-08-05T15:54:20.894403Repositório 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 |
|
_version_ |
1808128581714313216 |