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分别使用QuickGene-810型自动核酸提取仪法和手工试剂盒法,提取0.1%~100%梯度的转基因大豆GTS-40-3-2和非转基因大豆的DNA。使用核酸蛋白分析仪检测DNA浓度和纯度,荧光定量PCR扩增大豆内源Lectin基因和转基因大豆GTS-40-3-2结构特异性基因,对比QuickGene-810型自动核酸提取仪与手工方法的DNA提取效果。并用QuickGene-810型自动核酸提取仪法提取100份大豆样本DNA进行稳定性评价。结果表明:自动核酸提取仪法耗时短、成功率较高、稳定性良好,所得DNA的浓度和A260/A280比值均明显高于手工试剂盒法(P<0.001),荧光定量PCR扩增大豆内源Lectin基因的Ct值高于手工试剂盒法(P<0.001),但转基因大豆GTS-40-3-2结构特异性基因Ct值差别不明显(P=0.540)。
The DNA of transgenic soybean GTS-40-3-2 and non-transgenic soybean were extracted from 0.1% to 100% gradient by using QuickGene-810 automatic nucleic acid extraction method and manual kit method respectively. The nucleic acid protein analyzer was used to detect the concentration and purity of DNA. Fluorescence quantitative PCR was used to amplify the endogenous Lectin gene of soybean and GTS-40-3-2 gene of transgenic soybean. Compared with DNA of QuickGene-810 automatic nucleic acid extraction instrument and manual method Extract the effect. 100 samples of soybean samples were extracted with the QuickGene-810 automatic nucleic acid extraction system for stability evaluation. The results showed that the method of automatic nucleic acid extraction was shorter, the success rate was higher, and the stability was good. The concentration of DNA and the ratio of A260 / A280 were significantly higher than that of manual kit (P <0.001) The Ct value of the endogenous Lectin gene was higher than that of the manual kit method (P <0.001), but the Ct value of the GTS-40-3-2 structure-specific gene of transgenic soybean was not significantly different (P = 0.540).