Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum Biological Sciences |
Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689 |
Resumo: | DNA extraction methods were evaluated for the yield and purity of DNA recovered from mycorrhized roots and whether the recovered DNA is suitable for amplification of arbuscular mycorrhizal (AM) fungal SSU rDNA. The DNeasy Plant Mini Kit and three extraction buffers were used alone or in combination with either polyvinylpyrrolidone (PVP), polyvinylpolypyrrolidone (PVPP) and/or activated charcoal (AC). Among the extraction methods tested, those based on the CTAB buffers yielded more DNA than those based on the TE buffer and the DNeasy Plant Mini Kit. Moreover, the use of AC alone or in combination with PVPP reduced DNA yield, while it significantly improved the purity of recovered DNA, whatever the extraction buffer. On the other hand, the success of nested-PCR amplification was negatively correlated with the amount of template DNA and positively correlated with the purity of recovered DNA. Three methods based on the TE buffer, two on the CTAB-βM buffer and one on the DNeasy Plant Mini Kit produced high-quality DNA in terms of purity and PCR performance. However, the TE buffer-based methods are less time consuming than the CTAB-βM buffer-based methods, and cheaper than the method based on the DNeasy Plant Mini Kit. |
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Acta Scientiarum Biological Sciences |
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Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCRactivated charcoalTE bufferDNA qualityDNA extraction methods were evaluated for the yield and purity of DNA recovered from mycorrhized roots and whether the recovered DNA is suitable for amplification of arbuscular mycorrhizal (AM) fungal SSU rDNA. The DNeasy Plant Mini Kit and three extraction buffers were used alone or in combination with either polyvinylpyrrolidone (PVP), polyvinylpolypyrrolidone (PVPP) and/or activated charcoal (AC). Among the extraction methods tested, those based on the CTAB buffers yielded more DNA than those based on the TE buffer and the DNeasy Plant Mini Kit. Moreover, the use of AC alone or in combination with PVPP reduced DNA yield, while it significantly improved the purity of recovered DNA, whatever the extraction buffer. On the other hand, the success of nested-PCR amplification was negatively correlated with the amount of template DNA and positively correlated with the purity of recovered DNA. Three methods based on the TE buffer, two on the CTAB-βM buffer and one on the DNeasy Plant Mini Kit produced high-quality DNA in terms of purity and PCR performance. However, the TE buffer-based methods are less time consuming than the CTAB-βM buffer-based methods, and cheaper than the method based on the DNeasy Plant Mini Kit.Universidade Estadual De Maringá2014-10-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/2168910.4025/actascibiolsci.v36i4.21689Acta Scientiarum. Biological Sciences; Vol 36 No 4 (2014); 433-441Acta Scientiarum. Biological Sciences; v. 36 n. 4 (2014); 433-4411807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689/13610Diédhiou, Abdala GambyBorges, Wardsson LustrinoSadio, Oumarde Faria, Sergio Mianainfo:eu-repo/semantics/openAccess2014-10-02T17:25:25Zoai:periodicos.uem.br/ojs:article/21689Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSciPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2014-10-02T17:25:25Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
title |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
spellingShingle |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR Diédhiou, Abdala Gamby activated charcoal TE buffer DNA quality |
title_short |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
title_full |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
title_fullStr |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
title_full_unstemmed |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
title_sort |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR |
author |
Diédhiou, Abdala Gamby |
author_facet |
Diédhiou, Abdala Gamby Borges, Wardsson Lustrino Sadio, Oumar de Faria, Sergio Miana |
author_role |
author |
author2 |
Borges, Wardsson Lustrino Sadio, Oumar de Faria, Sergio Miana |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Diédhiou, Abdala Gamby Borges, Wardsson Lustrino Sadio, Oumar de Faria, Sergio Miana |
dc.subject.por.fl_str_mv |
activated charcoal TE buffer DNA quality |
topic |
activated charcoal TE buffer DNA quality |
description |
DNA extraction methods were evaluated for the yield and purity of DNA recovered from mycorrhized roots and whether the recovered DNA is suitable for amplification of arbuscular mycorrhizal (AM) fungal SSU rDNA. The DNeasy Plant Mini Kit and three extraction buffers were used alone or in combination with either polyvinylpyrrolidone (PVP), polyvinylpolypyrrolidone (PVPP) and/or activated charcoal (AC). Among the extraction methods tested, those based on the CTAB buffers yielded more DNA than those based on the TE buffer and the DNeasy Plant Mini Kit. Moreover, the use of AC alone or in combination with PVPP reduced DNA yield, while it significantly improved the purity of recovered DNA, whatever the extraction buffer. On the other hand, the success of nested-PCR amplification was negatively correlated with the amount of template DNA and positively correlated with the purity of recovered DNA. Three methods based on the TE buffer, two on the CTAB-βM buffer and one on the DNeasy Plant Mini Kit produced high-quality DNA in terms of purity and PCR performance. However, the TE buffer-based methods are less time consuming than the CTAB-βM buffer-based methods, and cheaper than the method based on the DNeasy Plant Mini Kit. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-10-03 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689 10.4025/actascibiolsci.v36i4.21689 |
url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689 |
identifier_str_mv |
10.4025/actascibiolsci.v36i4.21689 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689/13610 |
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 |
Universidade Estadual De Maringá |
publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 36 No 4 (2014); 433-441 Acta Scientiarum. Biological Sciences; v. 36 n. 4 (2014); 433-441 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum Biological Sciences |
collection |
Acta Scientiarum Biological Sciences |
repository.name.fl_str_mv |
Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
||actabiol@uem.br |
_version_ |
1799317395981991936 |