Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA?
Autor(a) principal: | |
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Data de Publicação: | 2018 |
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/10198/15054 |
Resumo: | The aim of this work was to evaluate a fungal DNA extraction procedure with the lowest inputs in terms of time as well as of expensive and toxic chemicals, but able to consistently produce genomic DNA of good quality for PCR purposes. Two types of fungal biological material were tested - mycelium and conidia - combined with two protocols for DNA extraction using Sodium Dodecyl Sulphate (SDS) and Cetyl Trimethyl Ammonium Bromide as extraction buffers and glass beads for mechanical disruption of cell walls. Our results showed that conidia and SDS buffer was the combination that lead to the best DNA quality and yield, with the lowest variation between samples. This study clearly demonstrates that it is possible to obtain high yield and pure DNA from pigmented conidia without the use of strong cell disrupting procedures and of toxic reagents. Significance and Impact of the Study: There are numerous methods f or DNA extraction from fungi. Some rely on expensive commercial kits and/or equipments, unavailable for many laboratories, or make use of toxic chemicals such as chloroform, phenol and mercaptoethanol. This study clearly demonstrates that it is possible to obtain high yields of pure DNA from pigmented conidia without the use of strong and expensive cell disrupting procedures and of toxic reagents. The method herein described is simultaneously inexpensive and adequate to DNA extraction from several different types of fungi. |
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Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA?AspergillusDNA extractionFungiGenomicsPCRThe aim of this work was to evaluate a fungal DNA extraction procedure with the lowest inputs in terms of time as well as of expensive and toxic chemicals, but able to consistently produce genomic DNA of good quality for PCR purposes. Two types of fungal biological material were tested - mycelium and conidia - combined with two protocols for DNA extraction using Sodium Dodecyl Sulphate (SDS) and Cetyl Trimethyl Ammonium Bromide as extraction buffers and glass beads for mechanical disruption of cell walls. Our results showed that conidia and SDS buffer was the combination that lead to the best DNA quality and yield, with the lowest variation between samples. This study clearly demonstrates that it is possible to obtain high yield and pure DNA from pigmented conidia without the use of strong cell disrupting procedures and of toxic reagents. Significance and Impact of the Study: There are numerous methods f or DNA extraction from fungi. Some rely on expensive commercial kits and/or equipments, unavailable for many laboratories, or make use of toxic chemicals such as chloroform, phenol and mercaptoethanol. This study clearly demonstrates that it is possible to obtain high yields of pure DNA from pigmented conidia without the use of strong and expensive cell disrupting procedures and of toxic reagents. The method herein described is simultaneously inexpensive and adequate to DNA extraction from several different types of fungi.Biblioteca Digital do IPBRodrigues, PaulaVenâncio, ArmandoLima, Nelson2018-01-25T10:00:00Z20182018-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10198/15054engRodrigues, Paula; Venâncio, Armando; Lima, Nelson (2018). Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? Letters in Applied Microbiology. ISSN 0266-8254. 66, p. 32-370266-825410.1111/lam.12822info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-21T10:35:33Zoai:bibliotecadigital.ipb.pt:10198/15054Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:04:51.228064Repositório 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 |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
title |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
spellingShingle |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? Rodrigues, Paula Aspergillus DNA extraction Fungi Genomics PCR |
title_short |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
title_full |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
title_fullStr |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
title_full_unstemmed |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
title_sort |
Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? |
author |
Rodrigues, Paula |
author_facet |
Rodrigues, Paula Venâncio, Armando Lima, Nelson |
author_role |
author |
author2 |
Venâncio, Armando Lima, Nelson |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Biblioteca Digital do IPB |
dc.contributor.author.fl_str_mv |
Rodrigues, Paula Venâncio, Armando Lima, Nelson |
dc.subject.por.fl_str_mv |
Aspergillus DNA extraction Fungi Genomics PCR |
topic |
Aspergillus DNA extraction Fungi Genomics PCR |
description |
The aim of this work was to evaluate a fungal DNA extraction procedure with the lowest inputs in terms of time as well as of expensive and toxic chemicals, but able to consistently produce genomic DNA of good quality for PCR purposes. Two types of fungal biological material were tested - mycelium and conidia - combined with two protocols for DNA extraction using Sodium Dodecyl Sulphate (SDS) and Cetyl Trimethyl Ammonium Bromide as extraction buffers and glass beads for mechanical disruption of cell walls. Our results showed that conidia and SDS buffer was the combination that lead to the best DNA quality and yield, with the lowest variation between samples. This study clearly demonstrates that it is possible to obtain high yield and pure DNA from pigmented conidia without the use of strong cell disrupting procedures and of toxic reagents. Significance and Impact of the Study: There are numerous methods f or DNA extraction from fungi. Some rely on expensive commercial kits and/or equipments, unavailable for many laboratories, or make use of toxic chemicals such as chloroform, phenol and mercaptoethanol. This study clearly demonstrates that it is possible to obtain high yields of pure DNA from pigmented conidia without the use of strong and expensive cell disrupting procedures and of toxic reagents. The method herein described is simultaneously inexpensive and adequate to DNA extraction from several different types of fungi. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-01-25T10:00:00Z 2018 2018-01-01T00:00:00Z |
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/10198/15054 |
url |
http://hdl.handle.net/10198/15054 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Rodrigues, Paula; Venâncio, Armando; Lima, Nelson (2018). Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA? Letters in Applied Microbiology. ISSN 0266-8254. 66, p. 32-37 0266-8254 10.1111/lam.12822 |
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.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 |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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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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1799135296690847744 |