摘要
应用多重PCR结合变性高效液相色谱(DHPLC)技术,检测转基因番茄华番1号中的转基因成分。对番茄中特异的PG内源基因、华番1号中含有的CAMV35S、NOS和NPTII外源基因进行多重PCR扩增,将扩增产物测序验证。利用DH-PLC分离多重PCR产物,建立了多重PCR-DHPLC技术检测转基因番茄的方法;将样品进行稀释,确定了该方法的检测灵敏度。试验结果表明,所建立的多重PCR-DHPLC方法操作简便,快速准确,能够同时检测转基因番茄4个内、外源基因,检测限达0.5ng/μL,可用于番茄中转基因成分的检测。
Multiplex PCR-DHPLC method were used for detecting genetically modified components in Huafan NO.1 tomato.CAMV35S,NOS and NPTII genes were detected by multiplex PCR.The endogenesis PG gene was also be amplified.The multiplex PCR fragments were sequenced by company and separated by DHPLC.Various grads of samples were used for the sensitivity testing.The result showed that the multiplex PCR-DHPLC separation method was proposed for the simultaneous detection of 4 genes in Huafan NO.1 tomato.The limit of detection was 0.5ng/μL.This method was easy-done,rapid,accurate and highly efficient,which can be used for detecting transgenic tomato.
出处
《作物杂志》
CAS
CSCD
北大核心
2011年第2期28-31,共4页
Crops
基金
2011年国家质检总局项目
黑龙江省科技厅青年基金
编号:QC2010022
