基于UNK2基因的大豆种质资源普通PCR和实时荧光PCR检测方法

Studies on conventional PCR and real-time PCR method for detecting soybean germplasm resources based on UNK2 gene

根据一种新的大豆内源特异性基因(UNK2)序列设计合成引物和探针,建立了一种基于该基因的普通PCR和实时荧光PCR检测方法。结果显示,该方法在普通PCR和实时荧光PCR检测中均具有好的特异性(普通PCR扩增产物为660 bp),而在其他作物品种上没有扩增;灵敏度试验证明,普通PCR检测灵敏度为0.03 ng,实时荧光PCR检测灵敏度为3 pg。本研究建立的方法特异性好、灵敏度较高,非常适合各口岸实验室对进出境大豆种质资源进行快速分子鉴定。

According to a new reported soybean endogenous specific gene （UNK2） in literature, we established a conventional PCR and real-time PCR detection method by designing specific primers and probes.Results showed that the two methods are very specific in the soybean gerrnplasms, the conventional PCR produce a 660 bp fragment. Sensitivity test showed that the detection limit of conventional PCR was 0.03 ng, and that of real-time was 3 pg. The two methods are very useful for the people in the port laboratories to identify the soybean germplasm resources.