09/12/2024
Fake DNA in your qPCR? Comparative evaluation of leaching levels in PCR plates
Rafal Grzeskowiak, Eppendorf SE
Sandrine Hamels, Eric Gancarek, Eppendorf Application Technologies S.A.
This study provides a comparative analysis of UV-absorbing leachables and qPCR fluorescence signals of water samples cycled in PCR plates from several manufacturers. The observed leaching levels and resulting false DNA concentrations were considerably high and varied markedly in the majority of the non-Eppendorf plates tested. This indicates that leaching may strongly influence qPCR assays and interfere with both photometric
and fluorescence signal quantification, leading to poor reproducibility results.
Introduction
The laboratory plastics are an integral part of any PCR workflow and may have a direct and serious impact on experimental outcome and data validity. An increasing number of studies indicate that a large part of processing additives may be released (leach) from the consumable into the samples and pose a source of error in various assay systems including PCR.
The leaching effects are particularly relevant for plate-based assays, where a high variability in temperature conditions as well as position-dependent leaching can dramatically influence the data validity and reproducibility of a PCR/qPCR assay.