Modulation of DNA Polymerases with Gold Nanoparticles and their Applications in Hot‐Start PCR

Abstract

A new gold-nanoparticle (AuNP)-based strategy to dynamically modulate the activity of DNA polymerases and realize a hot-start (HS)-like effect in the polymerase chain reaction (PCR) is reported, which effectively prevents unwanted nonspecific amplification and improves the performance of PCRs. A high-fidelity Pfu DNA polymerase is employed as the model system. Interestingly, AuNPs inactivate the polymerase activity of Pfu at low temperature, thus resembling an antibody-based HS PCR. This inhibition effect of AuNPs is demonstrated for the preamplification polymerization activity of the PCR, which largely suppresses nonspecific amplification at temperatures between 30 and 60 degrees C and leads to highly specific and sensitive PCR amplification with Pfu. Significantly, the fidelity of Pfu is not sacrificed in the presence of AuNPs. Therefore, this AuNP-based HS strategy provides a straightforward and potentially versatile approach to realize high-performance PCR amplification.

CdTe quantum dots accelerate the speed of Pfu-based polymerase chain reaction

“…[21] In the present work, a novel fast PCR was achieved simply by modifying PCR cycling protocol (two-step PCR and shortened cycling times) on a standard thermal cycler in the presence of CdTe QDs based on Pfu polymerase. We found that the addition of CdTe QDs significantly reduced the time required.…”

Atomic Force Microscopy Study of the Interaction of DNA and Nanoparticles

“…Gold, silver, and carbon nanoparticles can enhance the specificity and sensitivity of the polymerase chain reaction (PCR), while at excess dosage these nanoparticles can inhibit DNA amplification [4,49,50]. Pre-occupation of DNA by NPs in the binding sites of proteins that are requisite for DNA replication, transcription, and repair processes (e.g.…”

Sensitive and specific colorimetric DNA detection by invasive reaction coupled with nicking endonuclease-assisted nanoparticles amplification

“…However, the concentration of DNA in biological samples is very low, thus, many DNA amplification methods have been developed, such as polymerase chain reaction (PCR), (Mi et al, 2009;Mullis and Faloona, 1987;Pollet et al, 2011) loop-mediated isothermal amplification (LAMP), (Hsieh et al, 2012;Notomi, 2000;Wong et al, 2014) rolling cycle amplification (RCA). (Cheng et al, 2009;Lizardi et al, 1998;Zhao et al, 2013) However, these methods are based on template amplification, which usually causes amplicon crosscontamination, and leads to false positive results.…”