以16S rRNA基因为例，通过实时PCR定量分析单个引物-模板错配的有害影响。
Quantification of the detrimental effect of a single primer-template mismatch by real-time PCR using the 16S rRNA gene as an example.
作者信息
Bru D, Martin-Laurent F, Philippot L
机构信息
INRA, University of Burgundy, Soil and Environmental Microbiology, CMSE, 17 Rue Sully, B.P. 86510, 21065 Dijon Cedex, France.
出版信息
Appl Environ Microbiol. 2008 Mar;74(5):1660-3. doi: 10.1128/AEM.02403-07. Epub 2008 Jan 11.
Abstract
We investigated the effects of internal primer-template mismatches on the efficiency of PCR amplification using the 16S rRNA gene as the model template DNA. We observed that the presence of a single mismatch in the second half of the primer extension sequence can result in an underestimation of up to 1,000-fold of the gene copy number, depending on the primer and position of the mismatch.
摘要
我们以16S rRNA基因作为模型模板DNA，研究了内部引物-模板错配对PCR扩增效率的影响。我们观察到，根据错配的引物和位置，引物延伸序列后半部分存在单个错配可能导致基因拷贝数被低估高达1000倍。
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