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根据3种转基因棉花品系的边界序列设计品系检测引物,建立了一种特异性检测转基因棉花品系MON531、MON1445和MON15985的多重PCR-DHPLC方法。以20种不同的转基因及非转基因作物DNA验证该方法的特异性,结果只有MON531、MON15985和MON1445有特异的品系扩增片段峰,而其他转基因和非转基因作物无品系扩增片段峰,表明该方法特异性强。灵敏度实验结果表明3种转基因棉花的检测下限均为1 ng,灵敏度高。建立的方法可用于转基因棉花MON531、MON1445和MON15985品系及含有其成分产品的筛查或定性检测。
A multiplex PCR-DHPLC method was developed for the specific detection of transgenic cotton lines MON531, MON1445 and MON15985 based on the lineage primers of the three transgenic cotton lines. The specificity of this method was validated using 20 different transgenic and non-transgenic crop DNA. As a result, only MON531, MON15985, and MON1445 had unique lineage amplified fragments, while the other non-lineage amplified fragments of transgenic and non-transgenic crops showed that Method specificity is strong. Sensitivity experiment results show that the detection limit of the three kinds of transgenic cotton is 1 ng and the sensitivity is high. The established method can be used for screening or qualitative detection of GM cotton MON531, MON1445 and MON15985 lines and products containing their components.