Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies

Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape t...

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Autores principales:	Vasanthaiah,Hemanth K.N., Katam,Ramesh, Sheikh,Mehboob B
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	differential display RT-PCR grape RNA isolation RT-PCR subtractive hybridization Vitis
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000300005
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spelling	oai:scielo:S0717-345820080003000052009-01-30Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studiesVasanthaiah,Hemanth K.N.Katam,RameshSheikh,Mehboob B differential display RT-PCR grape RNA isolation RT-PCR subtractive hybridization Vitis Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.3 20082008-07-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000300005en10.4067/S0717-34582008000300005
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	differential display RT-PCR grape RNA isolation RT-PCR subtractive hybridization Vitis
spellingShingle	differential display RT-PCR grape RNA isolation RT-PCR subtractive hybridization Vitis Vasanthaiah,Hemanth K.N. Katam,Ramesh Sheikh,Mehboob B Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
description	Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.
author	Vasanthaiah,Hemanth K.N. Katam,Ramesh Sheikh,Mehboob B
author_facet	Vasanthaiah,Hemanth K.N. Katam,Ramesh Sheikh,Mehboob B
author_sort	Vasanthaiah,Hemanth K.N.
title	Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
title_short	Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
title_full	Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
title_fullStr	Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
title_full_unstemmed	Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
title_sort	efficient protocol for isolation of functional rna from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000300005
work_keys_str_mv	AT vasanthaiahhemanthkn efficientprotocolforisolationoffunctionalrnafromdifferentgrapetissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies AT katamramesh efficientprotocolforisolationoffunctionalrnafromdifferentgrapetissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies AT sheikhmehboobb efficientprotocolforisolationoffunctionalrnafromdifferentgrapetissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies
_version_	1718441786393755648

Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies

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