Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/89632
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Type: | Journal article |
Title: | Evaluation of four endogenous reference genes and their real-time PCR assays for common wheat quantification in GMOs detection |
Author: | Huang, H. Cheng, F. Wang, R. Zhang, D. Yang, L. |
Citation: | PLoS One, 2013; 8(9):e75850-1-e75850-9 |
Publisher: | Public Library of Science |
Issue Date: | 2013 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Forster, R. |
Statement of Responsibility: | Huali Huang, Fang Cheng, Ruoan Wang, Dabing Zhang, Litao Yang |
Abstract: | Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs) detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L.) DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat. |
Keywords: | Plants, Genetically Modified Triticum Starch Synthase Plant Proteins DNA Primers Reproducibility of Results Polymorphism, Single Nucleotide Genes, Plant Genetic Variation Real-Time Polymerase Chain Reaction |
Rights: | © 2013 Huang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
DOI: | 10.1371/journal.pone.0075850 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0075850 |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest 2 |
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hdl_89632.pdf | Published version | 733.75 kB | Adobe PDF | View/Open |
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