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RESEARCH PRODUCT
Quantification of the Detrimental Effect of a Single Primer-Template Mismatch by Real-Time PCR Using the 16S rRNA Gene as an Example
Laurent PhilippotFabrice Martin-laurentDavid Brusubject
Base Pair MismatchGene DosageBiologyPolymerase Chain ReactionApplied Microbiology and BiotechnologyPrimer extensionlaw.inventionDNA POLYMERASElawRNA Ribosomal 16SMethodsCopy-number variationGenePolymerase chain reactionDNA PrimersADN CIBLE[SDV.EE]Life Sciences [q-bio]/Ecology environmentGeneticsEcologyRibosomal RNA16S ribosomal RNACOPIE DE GENEReal-time polymerase chain reactionPseudomonas aeruginosaPrimer (molecular biology)Food ScienceBiotechnologydescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2008-01-11 | Applied and Environmental Microbiology |