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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 14, No. 1, 2011
Bioline Code: ej11012
Full paper language: English
Document type: Note
Document available free of charge

Electronic Journal of Biotechnology, Vol. 14, No. 1, 2011

 en Selection of reference genes for normalization of quantitative real-time PCR in cell cultures of Cyclamen persicum check for this species in other resources
Hoenemann, Claudia & Hohe, Annette

Abstract

As a prerequisite for gene expression analyses in cell cultures of the ornamental crop Cyclamen persicum check for this species in other resources 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.

Keywords
gene expression analysis, in vitro propagation, primer design, somatic embryogenesis.

 
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