活性污泥中氨氧化菌实时荧光定量PCR标准品制备方法的比较

发布时间：2018-04-08 16:24

本文选题：氨氧化菌　切入点：绝对定量　出处：《实验室研究与探索》2017年10期

【摘要】：为了快速准确地检测活性污泥中氨氧化菌(AOB)的拷贝数,探求一种新的快速简便的绝对定量标准品的制备方法。利用纯化的PCR产物和重组质粒标准品分别进行绝对定量,比较两者熔解曲线、扩增曲线、标准曲线和绝对拷贝数等方面的差异,分析两种标准品制备方法应用方面的优势。结果显示,两种标准品制备的熔解曲线都呈单一尖峰、扩增曲线指数期平行、标准曲线R299%,PCR产物作为标准品得到的AOB绝对拷贝数小于质粒标准品的结果(P0.05),表明纯化的PCR产物作为标准品较制备重组质粒标准品而言,操作过程更加简便、经济、对人员专业要求更低,在其纯度较高的情况下,具有和重组质粒标准品一样的应用效果。
[Abstract]:In order to detect the copy number of AOB in activated sludge quickly and accurately, a new rapid and simple preparation method of absolute quantitative standard was explored.Using the purified PCR product and the recombinant plasmid standard product for absolute quantification, the differences of fusion curve, amplification curve, standard curve and absolute copy number were compared, and the advantages of the two preparation methods were analyzed.The results showed that the melting curves of the two kinds of standard products showed a single peak, and the exponential periods of the amplification curves were parallel.The absolute copy number of AOB obtained by standard curve R299product was smaller than that of plasmid standard product (P0.05N), which indicated that the operation of purified PCR product was simpler and more economical than that of preparing recombinant plasmid standard product.It has the same application effect as the recombinant plasmid standard in the case of high purity.
【作者单位】：苏州科技大学江苏省环境科学与工程重点实验室环境科学与工程学院;
【基金】：国家研发计划项目(2016YFC0401103) 国家自然科学基金项目(51478284,51408387) 苏州市分离净化材料与技术重点实验室项目(SZS201512) 江苏省高等教育教改研究项目(2015JSJG248)
【分类号】：Q503;X703

【相似文献】