Analysis of some factors affecting to detect genetically modified ingredients by fluorescence real time PCR in rice noodle
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摘要: 为提高米制品中转基因成分实时荧光PCR检测的灵敏度与检出率,以及优化样品中转基因成分的检出效果,以添加不同转基因含量的米制品(米粉干)为模拟转基因样品,对影响检测效果的因素包括样品颗粒细度、DNA提取过程中样品在CTAB裂解缓冲液中温育时间等因素进行分析。结果显示,当样品颗粒细度>100目、CTAB温育时间达到8h条件下,样品DNA提取及荧光PCR检测结果最好;在最优化的条件组合下,样品转基因成分的检出限可达到0.001%转基因含量,是通常认为的荧光PCR检出限0.01%的10倍。Abstract: This paper studied the main factors which affected the detection of genetically modified components in deep maching rice noodle with real- time fluorescent PCR method, so as to improve the detection sensitivity and effect.Simulate rice noodle sample with different content of genetically modified components was made and major factors that mainly affect the test results were analyzed, including samples particle fineness, samples in CTAB buffer solution temperature bath time. Results showed that when the sample fineness was 100 mesh and CTAB buffer incubating time was 8h, the results of DNA extraction and PCR detection were the best. Under optimal combination of conditions, the sample GMO detection limit was 0.001% GM content, which was 10 times of usually fluorescent PCR detection limit which was 0.01%.
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