摘要
精炼大豆油中DNA的提取是转基因检测中的难点。实验以大豆油为研究对象,建立了一种有效的DNA提取新方法——冷冻干燥法。通过添加回收实验比较进口试剂盒方法和冷冻干燥法,发现冷冻干燥法提取的DNA浓度高、灵敏度高。采用冷冻干燥法从5种市售一级大豆油中提取DNA,进行了大豆内源Lectin基因、外源CaMV35S启动子、NOS终止子实时荧光PCR检测。结果表明,采用冷冻干燥法提取的大豆精炼油DNA适合PCR检测,可用于转基因检测。
DNA Isolation from Soybean Oil is a hurdle of GMO ( Genetically modified organisms) detection, In this research, a new method to isolate DNA from soybean refine oil was established--freeze-drying method. The freeze-drying method have been compared with commercial DNA extraction kits through spiked recovery , the result shows that the former method obtained DNA of high concentration and the former method had high sensitivity. Then the freeze-drying method was tested on five commercial soybean refine oils by detection of endogenous Lectin gene and foreign genes (CaMV35S, NOS) using Real-time PCR. Result shows that DNA purified from oil by freeze-drying method can be used for Real-time PCR analysis, and further for GMO detection.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2009年第12期155-159,共5页
Food and Fermentation Industries
基金
转基因重大专项课题(2008ZX08012-001)