An improved method for high-quality RNA isolation from needles of adult maritime pine trees

Detalhes bibliográficos
Autor(a) principal: Azevedo, Herlânder
Data de Publicação: 2003
Outros Autores: Neto, T. Lino, Tavares, R. M.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/2826
Resumo: Conventional RNA isolation methods optimised for pine seedlings have been shown to produce poor quality RNA when applied to needles of adult pine trees. We describe here a modified procedure to isolate high pure RNA from needles of thirty-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides and RNases. Major changes were the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by denaturing gel electrophoresis and by spectrophotometry (A260/A230 and A260/A280). Total RNA could be successfully used for poly(A)+-RNA isolation and cDNA library construction.
id RCAP_67dfdc5684520f0023dc8f3f7f3dfe93
oai_identifier_str oai:repositorium.sdum.uminho.pt:1822/2826
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling An improved method for high-quality RNA isolation from needles of adult maritime pine treescDNA library constructionMaritime pine needlesRNA isolationScience & TechnologyConventional RNA isolation methods optimised for pine seedlings have been shown to produce poor quality RNA when applied to needles of adult pine trees. We describe here a modified procedure to isolate high pure RNA from needles of thirty-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides and RNases. Major changes were the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by denaturing gel electrophoresis and by spectrophotometry (A260/A230 and A260/A280). Total RNA could be successfully used for poly(A)+-RNA isolation and cDNA library construction.Fundação para a Ciência e a Tecnologia (FCT) - grant ref. SFRH/BD/3194/2000.International Society for Plant Molecular BiologyUniversidade do MinhoAzevedo, HerlânderNeto, T. LinoTavares, R. M.20032003-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/2826engAzevedo, H., Lino-Neto, T. & Tavares, R.M. An improved method for high-quality RNA isolation from needles of adult maritime pine trees. Plant Mol Biol Rep 21, 333–338 (2003). https://doi.org/10.1007/BF027725820735-964010.1007/BF02772582https://link.springer.com/article/10.1007/BF02772582info: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:RCAAP2023-07-21T12:35:29Zoai:repositorium.sdum.uminho.pt:1822/2826Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:31:20.974130Repositório 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 An improved method for high-quality RNA isolation from needles of adult maritime pine trees
title An improved method for high-quality RNA isolation from needles of adult maritime pine trees
spellingShingle An improved method for high-quality RNA isolation from needles of adult maritime pine trees
Azevedo, Herlânder
cDNA library construction
Maritime pine needles
RNA isolation
Science & Technology
title_short An improved method for high-quality RNA isolation from needles of adult maritime pine trees
title_full An improved method for high-quality RNA isolation from needles of adult maritime pine trees
title_fullStr An improved method for high-quality RNA isolation from needles of adult maritime pine trees
title_full_unstemmed An improved method for high-quality RNA isolation from needles of adult maritime pine trees
title_sort An improved method for high-quality RNA isolation from needles of adult maritime pine trees
author Azevedo, Herlânder
author_facet Azevedo, Herlânder
Neto, T. Lino
Tavares, R. M.
author_role author
author2 Neto, T. Lino
Tavares, R. M.
author2_role author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Azevedo, Herlânder
Neto, T. Lino
Tavares, R. M.
dc.subject.por.fl_str_mv cDNA library construction
Maritime pine needles
RNA isolation
Science & Technology
topic cDNA library construction
Maritime pine needles
RNA isolation
Science & Technology
description Conventional RNA isolation methods optimised for pine seedlings have been shown to produce poor quality RNA when applied to needles of adult pine trees. We describe here a modified procedure to isolate high pure RNA from needles of thirty-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides and RNases. Major changes were the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by denaturing gel electrophoresis and by spectrophotometry (A260/A230 and A260/A280). Total RNA could be successfully used for poly(A)+-RNA isolation and cDNA library construction.
publishDate 2003
dc.date.none.fl_str_mv 2003
2003-01-01T00:00:00Z
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/1822/2826
url http://hdl.handle.net/1822/2826
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Azevedo, H., Lino-Neto, T. & Tavares, R.M. An improved method for high-quality RNA isolation from needles of adult maritime pine trees. Plant Mol Biol Rep 21, 333–338 (2003). https://doi.org/10.1007/BF02772582
0735-9640
10.1007/BF02772582
https://link.springer.com/article/10.1007/BF02772582
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 International Society for Plant Molecular Biology
publisher.none.fl_str_mv International Society for Plant Molecular Biology
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
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection 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
_version_ 1799132821780955136

Registros relacionados