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当前位置: 首页 > 文章 > 多聚酶对转基因产品PCR检测的影响分析 中国油料作物学报 2010,32 (1) 110-114
Position: Home > Articles > Comparison of different Taq DNA polymerase in PCR amplification during GMO detection Chinese Journal of Oil Crop Sciences 2010,32 (1) 110-114

多聚酶对转基因产品PCR检测的影响分析

作  者：
夏玉平;吴刚;武玉花;张丽;卢长明
单  位：
中国农业科学院油料作物研究所
关键词：
转基因检测;定性PCR;CaMV35S启动子;非特异扩增;灵敏度
摘  要：
本文参考出入境检验检疫行业标准中转基因油菜外源基因的PCR检测体系,分析了18种不同DNA聚合酶对定性PCR检测转基因油菜OXY235中的CaMV35S启动子的影响。结果表明,不同DNA聚合酶对CaMV35S启动子定性PCR检测中非特异扩增的产生以及检测灵敏度影响较大。其中热启动Taq酶效果较好,目标条带清晰,检测灵敏度高,而且完全没有非特异性扩增条带,非常适合用于定性PCR检测。对于同样条件下无法有效扩增0.1%DNA样品的Taq酶,则不推荐用于检测途径。
译  名：
Comparison of different Taq DNA polymerase in PCR amplification during GMO detection
作  者：
XIA Yu-ping1,WU Gang1,WU Yu-hua1,ZHANG Li1,2,LU Chang-ming1(1.Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture,Wuhan 430062,China;2. College of Life Science,South-Central University for Nationalities,Wuhan 430073,China)
关键词：
GMO detection; Qualitative PCR; CaMV35S promoter; Non-specific amplification; Sensitivity
摘  要：
In GMO (genetically modified organism) detection process,PCR results are usually different among various laboratories. Non-specific amplification bands,primer-dimer and different detection sensitivity often occur. Taq DNA polymerase might be one of the reasons resulting in different PCR amplification. According to genetically modified rapeseed detection standard developed by Entry-exit Inspection & Quarantine of P. R.China,18 Taq DNA polymerase were analyzed to amplify CaMV35S promoter in transgenic rapeseed OXY235. The results showed that Taq polymerase has severe impact on non-specific amplification and detection sensitivity. Hot-Start Taq polymerase was better for qualitative PCR detection,which produced clear target band in good sensitivity without non-specific amplification. However,it was found that,if Taq polymerase could not effectively amplify 0.1% DNA samples,it was not recommended in qualitative PCR detection.

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