Abstract

PCR has been identified as one of the most important tools in molecular biology and clinical medicine. Improvement of the specificity and efficiency of PCR is often required, and it still remains a great challenge. Here, we introduce the use of dendrimer-entrapped gold nanoparticles (Au DENPs) with different terminal groups prepared using poly(amidoamine) (PAMAM) dendrimers of generation 5 (G5) as a novel class of enhancers to improve the specificity and efficiency of PCR amplification. We show that the optimum concentrations of all the tested Au DENPs are lower than those of the corresponding PAMAM dendrimers without gold nanoparticles (AuNPs). For amine-terminated [(Au(0))(51.2) -G5.NH(2)] DENPs, the optimum required concentration is slightly lower than that of G5.NH(2) dendrimers, whereas for glycidol-modified [(Au(0))(51.2) -G5.NGlyOH] and acetylated [(Au(0))(51.2) -G5.NHAc] DENPs, the optimum required concentrations are one and three magnitude lower than the corresponding dendrimers, respectively. Our results suggest that the entrapment of AuNPs within the dendrimer interior helps to reserve the 3D spherical morphology of dendrimers, allowing for enhanced interaction with the PCR components. Simultaneously, because of the existence of thermal conductive AuNPs, the enhanced local heat transfer rate may afford decreased chances of mispairing between primers and templates, which is beneficial for enhancing the PCR specificity and efficiency. Therefore, the use of Au DENPs as a novel class of PCR enhancers may enable both improved interaction with the PCR components and the thermal conductivity, which allow them to be used for enhancing different error-prone PCR systems.

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