ZHOU Guang-biao,CAI Ying,CHEN Wen-wan,et al.Application of Quick Gene810 Automated Nucleic Acid Extraction Instrument on Detection of Genetically Modified Soybean[J].Soybean Science,2014,33(03):434-437.[doi:10.11861/j.issn.1000-9841.2014.03.0434]
QuickGene-810型自动核酸提取仪在转基因大豆检测中的应用研究
- Title:
- Application of Quick Gene810 Automated Nucleic Acid Extraction Instrument on Detection of Genetically Modified Soybean
- 分类号:
- Q943.2
- 文献标志码:
- A
- 摘要:
- 分别使用QuickGene810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）。
- Abstract:
- To compare the DNA extraction effect between the two following methods,DNA was extracted from 0.1%-100% proportion gradient samples of genetically modified soybean GTS-40-3-2 and non GMO soybean,using QuickGene-810 automated nucleic acid extraction instrument method and manual DNA extraction kit method respectively.DNA concentration and purity were detected by the protein nucleic acid analyzer,and soybean endogenous Lectin gene and genetically modified soybean GTS-40-3-2 structurespecific gene were quantitatively amplificated.In addition,DNA was extracted from 100 soybean samples employing QuickGene810 automated nucleic acid extraction instrument method,so as to evaluate the stability of this method.Results indicated that,QuickGene-810 automated nucleic acid extraction instrument method took a short time,had a high success rate and good stability.The concentration and the A260/A280ratio of DNA extracted by QuickGene-810 automated nucleic acid extraction instrument method were significantly higher than the manual kit method(P＜0.001).For soybean endogenous Lectin gene,the Ct value of nucleic acid extraction instrument method was higher than that of manual kit method(P＜0.001).However,for genetically modified soybean GTS-40-3-2 structure specific gene,the Ct values of the two methods had no significant difference(P=0.540).
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备注/Memo
收稿日期：2013-10-25