摘要
为提高米制品中转基因成分实时荧光聚合酶链式反应检测的灵敏度与检出率,应用模拟转基因白粿干样品,对影响检测结果的主要因素,包括样品颗粒细度、DNA提取过程中样品在十六烷基三甲基溴化铵裂解缓冲液中温育时间等因素进行优化。结果表明:提取的DNA量在样品颗粒细度与十六烷基三甲基溴化铵缓冲液温育时间的9种处理组合间都有极显著差异,其中最优条件为样品颗粒细度>100目、十六烷基三甲基溴化铵缓冲液温育时间>8 h(过夜)。采用最优前处理组合,样品的主要转基因成分检出限从公认的转基因含量0.01%(m/m)降到0.001%(m/m),检测灵敏度提高10倍,有效提高了米制品转基因成分检测结果的准确性。
In order to improve the sensitivity and detection limits of transgenic components in rice products, pretreatment conditions for DNA extraction including sample particle size and hexadecyltrimethylammonium bromide （CTAB） lysis buffer were optimized using simulated transgenic rice cake samples. The results showed that the content of DNA had significant.differences among 9 treatment combinations of sample particle size and CTAB lysis buffer incubation time. The content of DNA extracted by the combination of sample particle size larger than 100 mesh and CTAB lysis buffer incubation time longer than 8 h （overnight） was the highest. The detection limit of major transgenic components in rice cake samples was reduced from transgenic content of 0.01% （m/m） to 0.001% （m/m）, suggesting that the sensitivity of fluorescence polymerase chain reaction （PCR） was improved by 10 times using the optimal pretreatment. The detection accuracy of genetically modified components in rice products was effectively improved as well.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2014年第2期243-247,共5页
Food Science
基金
福建出入境检验检疫局科技计划项目(FK2010-28
FK2013-02)
关键词
米制品(白粿干)
转基因成分
实时荧光聚合酶链反应
检出限
rice products （rice cake）
genetically modified ingredients
fluorescence real time-polymerase chain reaction（RT-PCR）
limit of detection
