CAJ | 학술논문

采用改良环介导等温扩增(LAMP)技术,建立一套准确、快速、可靠的用于转基因大豆检测及DNA提取的方法.以CP4-EPSPS外源基因为检测的目的片段,设计内、外引物和环引物,并采用改进的方法提取DNA,进行LAMP扩增,实际检测已知转基因大豆和市售大豆,通过肉眼观察白色沉淀,判断检测结果.改进的DNA提取方法与经典的CTAB方法相比,提取时间至少缩短了1 h,降低了提取成本.环介导等温扩增(LAMP)技术对转基因大豆的检出限为0.01%,是普通PCR方法的20倍.因此,改良的LAMP检测转基因大豆方法灵敏度高,耗时短,方法简便.
채용개량배개도등온확증(LAMP)기술,건립일투준학、쾌속、가고적용우전기인대두검측급DNA제취적방법.이CP4-EPSPS외원기인위검측적목적편단,설계내、외인물화배인물,병채용개진적방법제취DNA,진행LAMP확증,실제검측이지전기인대두화시수대두,통과육안관찰백색침정,판단검측결과.개진적DNA제취방법여경전적CTAB방법상비,제취시간지소축단료1 h,강저료제취성본.배개도등온확증(LAMP)기술대전기인대두적검출한위0.01%,시보통PCR방법적20배.인차,개량적LAMP검측전기인대두방법령민도고,모시단,방법간편.
A loop-mediated isothermal amplification( LAMP) technology was established to detect transgenic soybean. The sequence of exogenous gene CP4 — EPSPS was used as target sequence, to design outer primers, inner primers and loop primers. The DNA extraction method was improved. Through visual observation of white precipitate to judge results of detection. The improved CTAB method isolated total DNA quickly and reducing DNA- extraction cost as compared with the traditional CTAB method. The detection limit of the genetically modified soybeans by LAMP technology was 0. 01% which was superior to PCR. Results indicate that LAMP can provide a sensitive, rapid yet simple test for the detection of CP4- EPSPS in transgenic soybean.