Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

RNA extraction from mangrove tissues can be difficult due to the presence of polyphenolic and polysaccharide compounds upon cell disruption. Besides, a successful RNA isolation from mangrove tissues sometimes can be strain and species specific. Two different methods were used to extract RNA from tis...

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Autores principales:	Rubio-Piña,Jorge A, Zapata-Pérez,Omar
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2011
Materias:	mangrove plant tissues polysaccharides RNA isolation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000500011
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spelling	oai:scielo:S0717-345820110005000112012-04-17Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plantsRubio-Piña,Jorge AZapata-Pérez,Omar mangrove plant tissues polysaccharides RNA isolation RNA extraction from mangrove tissues can be difficult due to the presence of polyphenolic and polysaccharide compounds upon cell disruption. Besides, a successful RNA isolation from mangrove tissues sometimes can be strain and species specific. Two different methods were used to extract RNA from tissues (stems, leaves and roots) of black mangrove (Avicennia germinans) and white mangrove (Laguncularia racemosa), collected from the mangrove area at Progreso, Yucatan, Mexico. An optimized and modified total RNA isolation method was developed for these recalcitrant species. The protocol is based on the CTAB method including β-mercaptoethanol and PVPP with sequential Phenol/Chloroform/isoamyl alcohol extractions to remove protein, and polyphenols, followed by two selective purifications with LiCl and sodium acetate to eliminate polysaccharides. Although, the introduced modifications are not new, their addition proved to be decisive for success in RNA isolation. This modified procedure resulted in high quantity and quality RNA. The RNA obtained is suitable for cDNA synthesis, RT-PCR experiments and the phytochelatin synthase 1 gene amplification.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.5 20112011-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000500011en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	mangrove plant tissues polysaccharides RNA isolation
spellingShingle	mangrove plant tissues polysaccharides RNA isolation Rubio-Piña,Jorge A Zapata-Pérez,Omar Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
description	RNA extraction from mangrove tissues can be difficult due to the presence of polyphenolic and polysaccharide compounds upon cell disruption. Besides, a successful RNA isolation from mangrove tissues sometimes can be strain and species specific. Two different methods were used to extract RNA from tissues (stems, leaves and roots) of black mangrove (Avicennia germinans) and white mangrove (Laguncularia racemosa), collected from the mangrove area at Progreso, Yucatan, Mexico. An optimized and modified total RNA isolation method was developed for these recalcitrant species. The protocol is based on the CTAB method including β-mercaptoethanol and PVPP with sequential Phenol/Chloroform/isoamyl alcohol extractions to remove protein, and polyphenols, followed by two selective purifications with LiCl and sodium acetate to eliminate polysaccharides. Although, the introduced modifications are not new, their addition proved to be decisive for success in RNA isolation. This modified procedure resulted in high quantity and quality RNA. The RNA obtained is suitable for cDNA synthesis, RT-PCR experiments and the phytochelatin synthase 1 gene amplification.
author	Rubio-Piña,Jorge A Zapata-Pérez,Omar
author_facet	Rubio-Piña,Jorge A Zapata-Pérez,Omar
author_sort	Rubio-Piña,Jorge A
title	Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
title_short	Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
title_full	Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
title_fullStr	Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
title_full_unstemmed	Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants
title_sort	isolation of total rna from tissues rich in polyphenols and polysaccharides of mangrove plants
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000500011
work_keys_str_mv	AT rubiopinajorgea isolationoftotalrnafromtissuesrichinpolyphenolsandpolysaccharidesofmangroveplants AT zapataperezomar isolationoftotalrnafromtissuesrichinpolyphenolsandpolysaccharidesofmangroveplants
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Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants

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