转基因苜蓿草J101品系特异性定性PCR检测方法的建立

摘要

目的：建立转基因苜蓿草J101品系特异性定性PCR检测方法。方法根据转基因苜蓿草品系J1015’端外源插入片段与苜蓿草基因组 DNA 之间的邻接区序列设计引物,建立了转基因苜蓿草 J101品系特异性定性PCR检测方法，并对本方法的特异性、灵敏度进行了测定。结果建立的检测方法特异于转基因苜蓿草J101检测,检测最低DNA 浓度为(1imit of det ection,LOD)为80 pg,相当于50拷贝转基因苜蓿草J101基因组DNA。结论本研究建立的转基因苜蓿草J101品系特异性定性PCR 检测方法特异性好,灵敏度高,能够快速、准确地对转基因苜蓿草J101进行检测分析。%Objective To establish a event-specific qualitative PCR detection method for genetically modified (GM) alfalfa events J101. Methods The specific primer pairs based on the 5’junction sequence spanning the alfalfa DNA and inserted fragment of J101 were designed and then the PCR detection system was established. The specificity and sensitivity were analyzed. Results The qualitative PCR method was specific for GM alfalfa J101 detection, the limit of detection (LOD) were 80 pg J101 genomic DNA or 50 copies of alfalfa J101 genomic DNA. Conclusion The established event-specific PCR method for GM alfalfa J101 detection has a high specificity and good sensitivity, and is suitable for detection of J101 samples quickly and accurately.