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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J101 5’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 detection,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 method for the qualitative and quantitative PCR detection of transgenic alfalfa J101 strain. Methods According to the sequence of the adjacent region between the 5 ’end of J101 transgenic alfalfa line and the genomic DNA of alfalfa, a specific qualitative PCR method was established for the detection of the transgenic alfalfa J101 strain. The specificity, sensitivity The measurement was carried out. Results The established detection method was specific to transgenic alfalfa J101. The detection limit of minimal DNA concentration (LOD) was 80 pg, equivalent to 50 copies of transgenic alfalfa J101 genomic DNA. Conclusion The specific qualitative PCR detection method of the transgenic alfalfa J101 strain established in this study has good specificity and high sensitivity, and can rapidly and accurately detect the transgenic alfalfa J101.