Quantification of the detrimental effect of a single primer-template mismatch by real-time PCR using the 16S rRNA gene as an example
Quantification of the detrimental effect of a single primer-template mismatch by real-time PCR using the 16S rRNA gene as an example
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DOI：
10.1128/aem.02403-07
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发表时间：
2008-03-01
影响因子：
4.4
通讯作者：
Philippot, L.
中科院分区：
文献类型：
--
作者：
Bru, D.;Martin-Laurent, F.;Philippot, L.
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，在引物延伸序列的后半部分存在单个错配可能导致基因拷贝数的低估高达1000倍，这取决于引物和错配的位置。
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.