| Registro completo |
| Provedor de dados: |
Biological Sciences
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| País: |
Brazil
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| Título: |
Assessment of DNA extraction methods for detection of arbuscular mycorrhizal fungi in plant roots by nested-PCR
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| Autores: |
Diédhiou, Abdala Gamby
Borges, Wardsson Lustrino
Sadio, Oumar
de Faria, Sergio Miana
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| Data: |
2014-10-03
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| Ano: |
2014
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| Palavras-chave: |
Activated charcoal
TE buffer
DNA quality
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| 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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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689
10.4025/actascibiolsci.v36i4.21689
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| Editor: |
Universidade Estadual De Maringá
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| Relação: |
http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/21689/13610
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| Formato: |
application/pdf
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| Fonte: |
Acta Scientiarum. Biological Sciences; v. 36, n. 4 (2014); 433-441
Acta Scientiarum. Biological Sciences; v. 36, n. 4 (2014); 433-441
1807-863X
1679-9283
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