【摘要】 为确定食品和饲料中的猪源性成分,本试验优化了PCR检测猪源性成分的反应体系。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;25m mol/L的MgCl2为0、2、4、6μl时;dNTP为0、0.1、0.2、0.4μl时,其他PCR反应因素为最大量的结果。结果表明:优化的猪源性成分PCR反应体系中Taq DNA聚合酶、25mmol/L MgCl2、dNTP适宜浓度分别为2U,2μl,0.4μl。优化后的反应体系对猪肉DNA最低检测浓度为0.04ng/μl。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。更多还原

【Abstract】 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 20 μl was used by this test. The PCR reaction system was optimized when the Taq DNA polymerase are 0 U、1 U、2 U、3 U,and the MgCl2 are 0 μl、2 μl、4 μl、6 μl,and the dNTP are 0 μl、0.1 μl、0.2 μl、0.4 μl 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 2 U,2 μl and 0.4 μl 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.更多还原