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根据转基因抗除草剂玉米DAS-40278-9外源基因的旁侧序列,采用ABI Primer Express 3.0设计引物和Taq Man探针,建立转基因抗除草剂玉米DAS-40278-9品系特异实时荧光定量PCR检测方法。采用该检测方法对DAS-40278-9玉米含量为1%(质量分数)的标准样品进行检测,结果显示,采用建立的方法获得的标准曲线,通过斜率、相关系数、扩增效率判断是可靠的,待测样品的定量检测结果为1.024%,非常接近真实值。本试验表明建立的转基因抗除草剂玉米DAS-40278-9品系特异定量PCR检测方法的特异性好,灵敏度和准确度高,值得推广应用。
Based on the flanking sequences of foreign gene of herbicide-tolerant corn DAS-40278-9, ABI Primer Express 3.0 primer and TaqMan probe were used to establish transgenic herbicide-resistant corn DAS-40278-9 strain-specific real-time fluorescence quantitative PCR method. Using this method to test the standard sample with 1% (mass fraction) DAS-40278-9 corn, the results showed that the standard curve obtained by the established method can be reliably judged by the slope, correlation coefficient and amplification efficiency The quantitative test result of the sample to be tested is 1.024%, which is very close to the true value. This experiment showed that the established method of DAS-40278-9 corn specific transgenic herbicide-tolerant quantitative PCR detection method has the advantages of good specificity, high sensitivity and high accuracy, and is worth popularizing and applying.