食用精炼大豆油DNA提取及单管巢式PCR检测

DNA extraction from soybean oil and detection of genetically-modified components by single-tube nested PCR

摘要: 设计了针对转基因Roundup Ready大豆外源基因扩增的内外2对引物,分别位于CaMV35s启动子,CTP基因,CP4EPSPS基因区域,并成功对精炼大豆油中的外源基因进行单管巢式PCR检测。结果表明,用改良试剂盒法和冷冻干燥法提取大豆油中的DNA均可以满足单管巢式PCR检测要求;单管巢式PCR扩增对内外引物的Tm值有特殊要求,外引物的退火温度高于内引物。本实验建立的精炼大豆油转基因成分的单管巢式PCR检测方法特异性强,灵敏度高且快速方便。

Abstract: The two pairs of primers were synthesized according to genetically- modified soybean, located in the CaMV35s promoter, CTP gene, CP4EPSPS gene region, and detection method of transgenic factors by single-tube nested PCR was established.It was proved that DNA extraction by improved reagent kit method or freeze drying technology from soybean oil could meet the detection of single-tube nested PCR, and for single-tube nested PCR outer primers annealing temperature must be higher than inner primers.