引物和温度对血豆腐制品中鸡源性成分定量测定的影响

Effect of Primers and Temperature on Quantitative Determination of Chicken-Derived Ingredients in Blood Curd

为了能够更加准确的进行基于RT-PCR技术的血豆腐制品中鸡源性成分定量检测,本实验从扩增DNA片段长度、标准样品DNA处理温度两方面对定量测定结果的影响进行研究。结果表明:高温灭菌会对DNA产生一定损伤,扩增DNA片段长度越短,越不容易受到高温对测定结果的影响,利用和待测样品灭菌温度条件相近的标准样品制作标准曲线,可以提高结果的准确性,完善血豆腐中鸡源性成分定量检测技术。

The effects of amplified fragment length and sterilization temperature of DNA standard sample on results of quantitative determination of chicken-derived ingredients in blood curd were investigated to establish an accurate quantitative RT-PCR assay. High-temperature sterilization induced DNA damage and PCR detection was less vulnerable to the effect of high-temperature sterilization when shorter DNA fragments were amplified. The accuracy of PCR detection was improved and as a result, a perfected PCR assay was established for quantitative determination of chicken-derived ingredients in blood curd when the standard sample used to prepare a standard curve was sterilized at a similar temperature as samples tested.