摘要
为建立一种可同步检测3种外源基因的多重PCR方法,试验选择较常见的T-CaMV 35S,P-CaMV 35S和pat这3个基因作为多重PCR检测对象,对多重PCR体系中引物浓度配比和引物退火温度进行优化。结果表明,多重PCR适宜的引物终浓度(μmol/L)配比T-CaMV 35S∶P-CaMV 35S∶pat为0.2∶0.2∶0.2以及适宜的引物退火温度为56℃;采用优化后的反应体系对多重PCR体系灵敏度进行测试,结果显示,方法灵敏度为0.5 ng;通过利用建立的多重PCR方法对已知样品进行检测,结果表明,试验建立的多重PCR体系具有良好的可靠性。
To establish a multiplex PCR method for detecting three kinds of exogenous genes, the T-CaMV 35 S, P-CaMV 35 S and pat were selected as parameters of multiple PCR detection. The reaction conditions were optimized. The experimental results showed that the optimum annealing temperature of multiple PCR was 56 ℃, the optimum final concentration of primers（μmol/L）ratio was 0.2∶0.2∶0.2 for T-CaMV 35S∶P-CaMV 35S∶pat. Using the optimized reaction system to test the sensitivity of multiplex PCR system, the results showed that the sensitivity of the method was 0.5 ng. Using the method of multiple PCR to detect the known samples, the test results showed that the multiplex PCR system was established with good reliability.
出处
《山西农业科学》
2017年第1期37-40,88,共5页
Journal of Shanxi Agricultural Sciences
基金
四川省财政现代农业技术创新与示范专项资金项目(2014CXSF-040)
四川省教育厅自然科学一般项目(15ZB0331)
作者简介
尹全（1981-），男，四川金堂人，助理研究员，硕士，主要从事转基因植物安全评价工作。
李忆为通信作者。
