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Comparison of Four Qualitative PCR-based Detection Methords of CaMV35S and tNOS Elements in Transgenic Plants |
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Abstract Promoter of Cauliflower mosaicvirus 35S(CaMV35S) and terminator of nopaline synthase gene(tNOS) are preferred parameters in screening detection of genetically modified organisms(GMO). In routine assay, some primers in stardands have non-specific amplification and low sensibility. In this study, 4 pairs of primer were collected, of which amplification product length are 195, 165, 147 and 123 bp, respectively, for CaMV35S, and 180, 172, 165 and 118 bp, for tNOS, respectively, and are used in different international and domestic standards. The specificity, sensitivity and amplification of 8 primers were tested and evaluated through ordinary PCR and Real-time PCR. The result showed that the primers with amplification product length of 165 and 147 bp, for CaMV35S, respectively, 172 and 165 bp, for tNOS, respectively, presented higher specificity and sensitivity, and were very suitable for GMO screening with high efficiency in detecting different processed products. However, CaMV35S 195 bp primer and tNOS 180 bp primer amplified weakly and nonspecificly, and that of CaMV35S 123 bp primer and tNOS 118 bp primer were weak and unstable. The study evaluate the primers in different international and domestic standards through ordinary PCR and Real-time PCR, and will be useful for the detection and supervision of GMOs.
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Received: 08 January 2014
Published: 02 February 2015
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