Modulation of DNA polymerases with gold nanoparticles and their applications in hot-start PCR.
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.
Mi
Lijuan
L
Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, P.R. China.
Wen
Yanqin
Y
Pan
Dun
D
Wang
Yanhong
Y
Fan
Chunhai
C
Hu
Jun
J
D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000648:ultrastructure*; D006046:Gold / Q000737:chemistry*; D008422:Materials Testing; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure*; D016133:Polymerase Chain Reaction / Q000379:methods*
D006046:Gold; D004259:DNA-Directed DNA Polymerase