A micromethod for high throughput RNA extraction in forest trees

A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA ex...

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Autores principales:	LE PROVOST,GREGOIRE, HERRERA,RAÚL, PAIVA,JORGE AP, CHAUMEIL,PHILIPPE, SALIN,FRANCK, PLOMION,CHRISTOPHE
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2007
Materias:	RNA extraction micromethod forest trees
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000400003
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spelling	oai:scielo:S0716-976020070004000032008-04-21A micromethod for high throughput RNA extraction in forest treesLE PROVOST,GREGOIREHERRERA,RAÚLPAIVA,JORGE APCHAUMEIL,PHILIPPESALIN,FRANCKPLOMION,CHRISTOPHE RNA extraction micromethod forest trees A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA extraction protocols are available but they are highly time consuming, and not adapted to large scale extraction. Here we present a high-throughput RNA extraction protocol. This protocol was adapted to a micro-scale by modifying the classical cetyltrimethylammonium (CTAB) protocol developed for pine: (i) quantity of material used (100-200 mg of sample), (ii) disruption of samples in microtube using a mechanical tissue disrupter, and (iii) the use of SSTE buffer. One hundred samples of woody plant tissues/organs can be easily treated in two working days. An average of 15 \ig of high quality RNA per sample was obtained. The RNA extracted is suitable for applications such as real time reverse transcription polymerase chain reaction, cDNA library construction or synthesis of complex targets for microarray analysisinfo:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.40 n.3 20072007-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000400003en10.4067/S0716-97602007000400003
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	RNA extraction micromethod forest trees
spellingShingle	RNA extraction micromethod forest trees LE PROVOST,GREGOIRE HERRERA,RAÚL PAIVA,JORGE AP CHAUMEIL,PHILIPPE SALIN,FRANCK PLOMION,CHRISTOPHE A micromethod for high throughput RNA extraction in forest trees
description	A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA extraction protocols are available but they are highly time consuming, and not adapted to large scale extraction. Here we present a high-throughput RNA extraction protocol. This protocol was adapted to a micro-scale by modifying the classical cetyltrimethylammonium (CTAB) protocol developed for pine: (i) quantity of material used (100-200 mg of sample), (ii) disruption of samples in microtube using a mechanical tissue disrupter, and (iii) the use of SSTE buffer. One hundred samples of woody plant tissues/organs can be easily treated in two working days. An average of 15 \ig of high quality RNA per sample was obtained. The RNA extracted is suitable for applications such as real time reverse transcription polymerase chain reaction, cDNA library construction or synthesis of complex targets for microarray analysis
author	LE PROVOST,GREGOIRE HERRERA,RAÚL PAIVA,JORGE AP CHAUMEIL,PHILIPPE SALIN,FRANCK PLOMION,CHRISTOPHE
author_facet	LE PROVOST,GREGOIRE HERRERA,RAÚL PAIVA,JORGE AP CHAUMEIL,PHILIPPE SALIN,FRANCK PLOMION,CHRISTOPHE
author_sort	LE PROVOST,GREGOIRE
title	A micromethod for high throughput RNA extraction in forest trees
title_short	A micromethod for high throughput RNA extraction in forest trees
title_full	A micromethod for high throughput RNA extraction in forest trees
title_fullStr	A micromethod for high throughput RNA extraction in forest trees
title_full_unstemmed	A micromethod for high throughput RNA extraction in forest trees
title_sort	micromethod for high throughput rna extraction in forest trees
publisher	Sociedad de Biología de Chile
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000400003
work_keys_str_mv	AT leprovostgregoire amicromethodforhighthroughputrnaextractioninforesttrees AT herreraraul amicromethodforhighthroughputrnaextractioninforesttrees AT paivajorgeap amicromethodforhighthroughputrnaextractioninforesttrees AT chaumeilphilippe amicromethodforhighthroughputrnaextractioninforesttrees AT salinfranck amicromethodforhighthroughputrnaextractioninforesttrees AT plomionchristophe amicromethodforhighthroughputrnaextractioninforesttrees AT leprovostgregoire micromethodforhighthroughputrnaextractioninforesttrees AT herreraraul micromethodforhighthroughputrnaextractioninforesttrees AT paivajorgeap micromethodforhighthroughputrnaextractioninforesttrees AT chaumeilphilippe micromethodforhighthroughputrnaextractioninforesttrees AT salinfranck micromethodforhighthroughputrnaextractioninforesttrees AT plomionchristophe micromethodforhighthroughputrnaextractioninforesttrees
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A micromethod for high throughput RNA extraction in forest trees

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