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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
agricultural and biological sciences
Development of real-time PCR method for the detection and the quantification of a new endogenous reference gene in sugar beet "Beta vulgaris L.": GMO application
Plant Cell Reports, Volume 32, No. 1, Year 2013
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Description
Key message: Here, we describe a new developed quantitative real-time PCR method for the detection and quantification of a new specific endogenous reference gene used in GMO analysis. The key requirement of this study was the identification of a new reference gene used for the differentiation of the four genomic sections of the sugar beet (Beta vulgaris L.) (Beta, Corrollinae, Nanae and Procumbentes) suitable for quantification of genetically modified sugar beet. A specific qualitative polymerase chain reaction (PCR) assay was designed to detect the sugar beet amplifying a region of the adenylate transporter (ant) gene only from the species of the genomic section I of the genus Beta (cultivated and wild relatives) and showing negative PCR results for 7 species of the 3 other sections, 8 related species and 20 non-sugar beet plants. The sensitivity of the assay was 15 haploid genome copies (HGC). A quantitative real-time polymerase chain reaction (QRT-PCR) assay was also performed, having high linearity (R2 > 0. 994) over sugar beet standard concentrations ranging from 20,000 to 10 HGC of the sugar beet DNA per PCR. The QRT-PCR assay described in this study was specific and more sensitive for sugar beet quantification compared to the validated test previously reported in the European Reference Laboratory. This assay is suitable for GMO quantification in routine analysis from a wide variety of matrices. © 2012 Springer-Verlag Berlin Heidelberg.
Authors & Co-Authors
Chaouachi, Maher
Tunisia, Monastir
Institut Supérieur de Biotechnologie de Monastir
Alaya, Akram
Tunisia, Monastir
Chu Fattouma-bourguiba
Ali, Imen Ben Haj
Tunisia, Tunis
University of Carthage, Institut National Des Sciences Appliquées et de Technologie
Ben Hafsa, Ahmed
Tunisia, Monastir
Institut Supérieur de Biotechnologie de Monastir
Nabi, Nesrine
Tunisia, Monastir
Institut Supérieur de Biotechnologie de Monastir
Bérard, Aurélie
France, Evry
Centre National de Recherche en Génomique Humaine
Romaniuk, Marcel
France, Versailles
Centre de Recherche Île-de-france - Versailles-grignon
Harzallah-Skhiri, Féthia
Tunisia, Monastir
Institut Supérieur de Biotechnologie de Monastir
Saïd, Khaled H.
Tunisia, Monastir
Institut Supérieur de Biotechnologie de Monastir
Statistics
Citations: 21
Authors: 9
Affiliations: 5
Identifiers
Doi:
10.1007/s00299-012-1346-5
ISSN:
07217714
Research Areas
Genetics And Genomics
Study Approach
Qualitative
Quantitative