Quantification of the detrimental effect of a single primer-template mismatch by real-time PCR using the 16S rRNA gene as an example

Applied and Environmental Microbiology
D BruL Philippot

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.

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Citations

Oct 12, 2010·Microbial Ecology·Laurent PhilippotEllen Kandeler
Jul 19, 2012·Microbial Ecology·Cristina Díez-VivesSilvia G Acinas
Jan 22, 2010·The ISME Journal·Anna EngelbrektsonPhilip Hugenholtz
Apr 12, 2012·Applied and Environmental Microbiology·Robert BrankatschkHelmut Bürgmann
May 5, 2012·BMC Microbiology·Dan-Ping MaoZhe-Xue Quan
Apr 17, 2013·Retrovirology·Alexander O PasternakBen Berkhout
Mar 9, 2013·Bioscience, Biotechnology, and Biochemistry·Sayed A M AmerMohammed Shobrak
Apr 27, 2012·PloS One·Ryuji J MachidaNancy Knowlton
Aug 1, 2012·PloS One·John Christian Gaby, Daniel H Buckley
Dec 10, 2013·Systematic and Applied Microbiology·Suvidha SamantDittmar Hahn
Jan 23, 2014·The Science of the Total Environment·Björn BerglundPer-Eric Lindgren
Sep 12, 2014·Biology Letters·Bruce E DeaglePierre Taberlet
Jan 14, 2010·Analytical and Bioanalytical Chemistry·Nina PapazovaIsabel Taverniers
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