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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。
The primers and probes were designed based on a novel sequence of endogenous specific gene (UNK2) of soybean, and a common PCR and real-time PCR detection method based on this gene was established. The results showed that this method has good specificity (both ordinary PCR and real-time fluorescence PCR detection of 660 bp), but not in other crop varieties; sensitivity test showed that the sensitivity of ordinary PCR detection was 0.03 ng, real-time fluorescence PCR detection sensitivity of 3 pg. The method established in this study has good specificity and high sensitivity, and is very suitable for rapid molecular identification of inbound and outbound soybean germplasm resources in various port labs.