抗草甘膦转基因大豆PCR-ELISA检测方法的建立

Yuan Qiang , Wei Yun-min , Fu Ming-ming , Qiu You-wen , Wen Hong-tao , Zhang Ming-hui , Liu Ying , Ao Jin-xia
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引用次数: 0

摘要

建立并优化了抗草甘膦转基因大豆的PCR-ELISA检测方法。结果显示,PCR-ELISA的关键参数如下:地高辛标记探针的浓度为0.5µmol·L−1,杂交反应时间为15 min,显色反应时间为30 min。对PCR-ELISA方法的灵敏度和重复性进行了评价,结果表明,当转基因大豆样品DNA模板的浓度为0.01%或更高时,该方法均可检测到,在我们的研究条件下,该方法的变异系数小于5%。上述结果表明,本研究建立的PCR-ELISA检测转基因大豆样品的方法重复性好,精密度高。
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Establishment of PCR-ELISA for Detecting Glyphosate Resistant Transgenic Soybean

A PCR-ELISA method for detecting the glyphosate resistant transgenic soybean was established and optimized. The results showed that the key parameters of PCR-ELISA were as follows: the concentration of digoxin tag probe was 0.5 µmol · L−1, the time of hybridization reaction was 15 min and the chromogenic reaction should last for 30 min. The sensitivity and the repeatability of our PCR-ELISA method were evaluated, and the results showed that it could be detected when the concentration of DNA template from transgenic soybean samples was 0.01% or higher, and the coefficient of variation of this method was less than 5% in our research condition. These results suggested that PCR-ELISA method establishment in this study had good repeatability and high precision for detecting the transgenic soybean samples.

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