猪源性成分的PCR检测技术优化研究

本试验研究目的是对食品和饲料中的猪源性成分PCR检测技术的反应体系进行优化。试验采用20μL的 PCR反应体系，分别测定Taq DNA聚合酶为0U、1U、2U、3U时；25mMol?L-1的MgCl2 为0μL、2μL、4μL、6μL时；dNTP为0μL、0.1μL、0.2μL、0.4μL时，其它PCR反应因素为最大量的结果，以优化猪源性成分PCR反应体系。试验结果：1、优化的猪源性成分PCR反应体系中Taq DNA聚合酶、25mMol?L-1 MgCl2、dNTP适宜浓度分别为2U，2μL，0.4μL。2、优化的猪源性成分PCR反应体系其检测最低猪肉DNA浓度为0.04 ng?μL-1。结论：本研究优化出的猪源性成分PCR反应体系经二重PCR反应技术进行验证，不仅快速，简便，准确性与灵敏度高，并且经济、高效。

Optimization of PCR system for pig component detection

The purpose of this study was to optimize the PCR reaction system of molecular technique for detecting the pig component in the food and feed. The PCR reaction system of 20μl was used by this test. The PCR reaction system was optimized when the Taq DNA polymerase are 0U, 1U,2U,3 U, and the MgCl2 are 0μl,2μll,4μll,6μll, and the dNTP are 0μl,0.1μl,0.2μl, 0.4μl and the other reaction factors of PCR reaction system are the maximum. The result is that ： the fitting concentrations for the Taq DNA polymerase,25 mMol/L MgCl2 and dNTP in the PCR reaction system are 2U, 2μl and 0.4μl respectively; 2, the minimum concentration of DNA of pig components in this optimizational PCR reaction system can be detected is 0.04 ng/μl.The conclusion： The optimal PCR reaction system for the pig component which has been validated by the Duplex PCR technique showes that it can work accurately and efficiently.