Establishment of Droplet Digital PCR Quantitative Detection Method of Transgenic Rice Strain‘Bt Shanyou 63’

doi:10.11924/j.issn.1000-6850.casb17030012

Abstract

Abstract: In order to improve the quantitative detection efficiency of transgenic rice, to promote the implement of transgenic food quantitative identification system, droplet digital PCR quantitative detection method of transgenic rice ‘Bt shanyou 63’ was studied in this paper. Based on the exogenous inserting sequence of transgenic rice ‘Bt shanyou 63’ and the rice sucrose phosphate synthase (SPS) gene, high efficiency primers and probes were selected, designed，and synthesized to amplify the endogenous gene and the strain specific sequence of ‘Bt shanyou 63’ with droplet digital PCR. The result of primers and probes optimizing experiment demonstrated that SPS60-F/R/P and ‘Bt shanyou 63’- F/R/P was suitable for dual channel ddPCR method to quantify transgenic rice ‘Bt shanyou 63’. The accuracy experiment showed that the measured value is similar to the standard value, the biases between the standard value and the measured value of 4 standard samples were 0.03, 0.15, 0.15 and 0.03. The SDs of measured value were 0.08-0.48, the RSDs were among 4.86% and 15.10%, less than the requirement of European Union, indicating that the accuracy was good. The established detection system of droplet ddPCR method for transgenic rice line ‘Bt 63’ quantification is simple, with high efficiency and accuracy, and can be used for quantitative analysis of transgenic rice line ‘Bt 63’ in agricultural products and foods.

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