Isolation of functional total RNA from Argemone mexicana tissues
RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and ther...
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Pontificia Universidad Católica de Valparaíso
2008
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oai:scielo:S0717-345820080004000152009-06-16Isolation of functional total RNA from Argemone mexicana tissuesRubio-Piña,Jorge AVázquez-Flota,Felipe A Argemone mexicana plant tissues RNA isolation secondary metabolites RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel electrophoresis. Moreover, the RNA obtained by this method, could be used directly for both RT-PCR and northern blot analysis, without any further purification.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.4 20082008-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000400015en10.4067/S0717-34582008000400015 |
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Scielo Chile |
collection |
Scielo Chile |
language |
English |
topic |
Argemone mexicana plant tissues RNA isolation secondary metabolites |
spellingShingle |
Argemone mexicana plant tissues RNA isolation secondary metabolites Rubio-Piña,Jorge A Vázquez-Flota,Felipe A Isolation of functional total RNA from Argemone mexicana tissues |
description |
RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel electrophoresis. Moreover, the RNA obtained by this method, could be used directly for both RT-PCR and northern blot analysis, without any further purification. |
author |
Rubio-Piña,Jorge A Vázquez-Flota,Felipe A |
author_facet |
Rubio-Piña,Jorge A Vázquez-Flota,Felipe A |
author_sort |
Rubio-Piña,Jorge A |
title |
Isolation of functional total RNA from Argemone mexicana tissues |
title_short |
Isolation of functional total RNA from Argemone mexicana tissues |
title_full |
Isolation of functional total RNA from Argemone mexicana tissues |
title_fullStr |
Isolation of functional total RNA from Argemone mexicana tissues |
title_full_unstemmed |
Isolation of functional total RNA from Argemone mexicana tissues |
title_sort |
isolation of functional total rna from argemone mexicana tissues |
publisher |
Pontificia Universidad Católica de Valparaíso |
publishDate |
2008 |
url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000400015 |
work_keys_str_mv |
AT rubiopinajorgea isolationoffunctionaltotalrnafromargemonemexicanatissues AT vazquezflotafelipea isolationoffunctionaltotalrnafromargemonemexicanatissues |
_version_ |
1718441792395804672 |