Department of Environmental Science and Engineering, Ewha Womans University, Seodaemun-gu, Seoul, Korea.
J Environ Sci Health A Tox Hazard Subst Environ Eng. 2013;48(4):417-21. doi: 10.1080/10934529.2013.728915.
Isolation of reference DNA templates for quantitative real-time PCR assays is an expensive, labor-intensive and time-consuming process if they are not readily available. Two artificial DNA templates with multiple probe sites were designed for quantifying methanogens and their 10 subgroups, based on the methyl coenzyme M reductase gene (mcrA). Their standards were comparable to each other. PCR amplification efficiencies (cycle vs. cumulative fluorescence) of the artificial DNAs were also comparable to those of the observed methanogen groups from anaerobic digesters. The artificial templates can be alternatives to the actual references.
如果定量实时 PCR 分析的参考 DNA 模板不易获得，那么其分离过程既昂贵又耗时，且需要大量人力。本研究基于甲基辅酶 M 还原酶基因（mcrA），设计了 2 个包含多个探针位点的人工 DNA 模板，用于定量检测产甲烷菌及其 10 个亚群。这两个标准品彼此相当。从厌氧消化池中观察到的产甲烷菌的 PCR 扩增效率（循环数与累积荧光强度）与人工 DNA 相当。人工模板可以作为实际参考的替代品。
