Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum

As a prerequisite for gene expression analyses in cell cultures of the ornamental crop Cyclamen persicum basic parameters for quantitative real-time polymerase chain reaction (qRT-PCR) have been established including the selection of reference genes using the software tools ‘geNorm&#82...

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Autores principales:	Hoenemann,Claudia, Hohe,Annette
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2011
Materias:	gene expression analysis in vitro propagation primer design somatic embryogenesis
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100012
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spelling	oai:scielo:S0717-345820110001000122011-05-24Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicumHoenemann,ClaudiaHohe,Annette gene expression analysis in vitro propagation primer design somatic embryogenesis As a prerequisite for gene expression analyses in cell cultures of the ornamental crop Cyclamen persicum basic parameters for quantitative real-time polymerase chain reaction (qRT-PCR) have been established including the selection of reference genes using the software tools ‘geNorm’ and ‘NormFinder’. Five potential reference genes have been tested (elongation factor tu (Ef-Tu), putative ABC transporter ATPase, putative conserved oligomeric Golgi (COG) complex component, V-ATPase G subunit 1 and Histone H3-K9 methyltransferase 4 (H3-K9-HMTase 4)). ‘NormFinder’ as well as ‘geNorm’ identified Ef-Tu to be the least stable reference gene while the ranking of the most stable genes differed depending on the algorithm. According to ‘NormFinder’ COG complex component displayed the most stable expression whereas ‘geNorm’ indicated V-ATPase G subunit 1 and a putative ABC transporter ATPase to be the most reliable reference genes. Hence, we concluded to use a normalization factor calculated from the four reference genes V-ATPase G subunit 1, ABC transporter ATPase, Histone H3-K9 methyltransferase 4 (H3-K9-HMTase 4) and COG complex component for normalization of qRT-PCR in cell cultures of Cyclamen persicum.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.1 20112011-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100012en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	gene expression analysis in vitro propagation primer design somatic embryogenesis
spellingShingle	gene expression analysis in vitro propagation primer design somatic embryogenesis Hoenemann,Claudia Hohe,Annette Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
description	As a prerequisite for gene expression analyses in cell cultures of the ornamental crop Cyclamen persicum basic parameters for quantitative real-time polymerase chain reaction (qRT-PCR) have been established including the selection of reference genes using the software tools ‘geNorm’ and ‘NormFinder’. Five potential reference genes have been tested (elongation factor tu (Ef-Tu), putative ABC transporter ATPase, putative conserved oligomeric Golgi (COG) complex component, V-ATPase G subunit 1 and Histone H3-K9 methyltransferase 4 (H3-K9-HMTase 4)). ‘NormFinder’ as well as ‘geNorm’ identified Ef-Tu to be the least stable reference gene while the ranking of the most stable genes differed depending on the algorithm. According to ‘NormFinder’ COG complex component displayed the most stable expression whereas ‘geNorm’ indicated V-ATPase G subunit 1 and a putative ABC transporter ATPase to be the most reliable reference genes. Hence, we concluded to use a normalization factor calculated from the four reference genes V-ATPase G subunit 1, ABC transporter ATPase, Histone H3-K9 methyltransferase 4 (H3-K9-HMTase 4) and COG complex component for normalization of qRT-PCR in cell cultures of Cyclamen persicum.
author	Hoenemann,Claudia Hohe,Annette
author_facet	Hoenemann,Claudia Hohe,Annette
author_sort	Hoenemann,Claudia
title	Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
title_short	Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
title_full	Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
title_fullStr	Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
title_full_unstemmed	Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum
title_sort	selection of reference genes for normalization of quantitative real-time pcr in cell cultures of cyclamen persicum
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2011
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000100012
work_keys_str_mv	AT hoenemannclaudia selectionofreferencegenesfornormalizationofquantitativerealtimepcrincellculturesofcyclamenpersicum AT hoheannette selectionofreferencegenesfornormalizationofquantitativerealtimepcrincellculturesofcyclamenpersicum
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Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum

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