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Identification and Quantification of Genetically Modified Moonshade Carnation Lines Using Conventional and TaqMan Real-Time Polymerase Chain Reaction Methods

文献类型： 外文期刊

作者： Li, Peng 1 ; Jia, Junwei 1 ; Bai, Lan 1 ; Pan, Aihu 1 ; Tang, Xueming 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotech Res Inst, Shanghai, Peoples R China

2.Shanghai Key Lab Agr Genet & Breeding, Shanghai, Peoples R China

关键词： Moonshade;Genetically modified organism;Conventional and TaqMan real-time PCR;Thermal asymmetric interlaced-PCR (TAIL-PCR)

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

ISSN： 0273-2289

年卷期： 2013 年 170 卷 5 期

页码：

收录情况： SCI

摘要： Genetically modified carnation (Dianthus caryophyllus L.) Moonshade was approved for planting and commercialization in several countries from 2004. Developing methods for analyzing Moonshade is necessary for implementing genetically modified organism labeling regulations. In this study, the 5'-transgene integration sequence was isolated using thermal asymmetric interlaced (TAIL)-PCR. Based upon the 5'-transgene integration sequence, conventional and TaqMan real-time PCR assays were established. The relative limit of detection for the conventional PCR assay was 0.05 % for Moonshade using 100 ng total carnation genomic DNA, corresponding to approximately 79 copies of the carnation haploid genome, and the limits of detection and quantification of the TaqMan real-time PCR assay were estimated to be 51 and 254 copies of haploid carnation genomic DNA, respectively. These results are useful for identifying and quantifying Moonshade and its derivatives.

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