Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient,...

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Autores principales:	Rai,Vineeta, Ghosh,Jayadri Sekhar, Dey,Nrisingha
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	acid phenol Bambusa balcooa fiber specific genes internode RNA extraction
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500022
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spelling	oai:scielo:S0717-345820100005000222011-05-24Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studiesRai,VineetaGhosh,Jayadri SekharDey,Nrisingha acid phenol Bambusa balcooa fiber specific genes internode RNA extraction RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be superior, when compared to total RNA extracted by other available protocols. The A260/A280 absorption ratio of the isolated RNA was found ranging between 1.89-1.97. The integrity of 28S and 18S rRNA was highly satisfactory when analyzed in agarose denaturing gel. RNA was further used for RT PCR, northern hybridization, cDNA library and subtractive hybridization without any further refinement.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.5 20102010-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500022en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	acid phenol Bambusa balcooa fiber specific genes internode RNA extraction
spellingShingle	acid phenol Bambusa balcooa fiber specific genes internode RNA extraction Rai,Vineeta Ghosh,Jayadri Sekhar Dey,Nrisingha Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
description	RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be superior, when compared to total RNA extracted by other available protocols. The A260/A280 absorption ratio of the isolated RNA was found ranging between 1.89-1.97. The integrity of 28S and 18S rRNA was highly satisfactory when analyzed in agarose denaturing gel. RNA was further used for RT PCR, northern hybridization, cDNA library and subtractive hybridization without any further refinement.
author	Rai,Vineeta Ghosh,Jayadri Sekhar Dey,Nrisingha
author_facet	Rai,Vineeta Ghosh,Jayadri Sekhar Dey,Nrisingha
author_sort	Rai,Vineeta
title	Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
title_short	Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
title_full	Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
title_fullStr	Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
title_full_unstemmed	Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
title_sort	isolation of total rna from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500022
work_keys_str_mv	AT raivineeta isolationoftotalrnafromhardbambootissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies AT ghoshjayadrisekhar isolationoftotalrnafromhardbambootissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies AT deynrisingha isolationoftotalrnafromhardbambootissuerichinpolyphenolsandpolysaccharidesforgeneexpressionstudies
_version_	1718441825327382528

Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies

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