Journal
SMALL
Volume 5, Issue 22, Pages 2597-2600Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/smll.200901147
Keywords
DNA; gold; nanoparticles; polymerase chain reaction
Categories
Funding
- National Natural Science Foundation [10674147, 20725516, 20873175]
- National Basic Research Program of China [2006CB933000, 2007CB936000, 2007AA06A406]
- Ministry of Health [2009ZX10004-301]
- Shanghai Municipal Commission for Science and Technology [0752nm021, 0952nm04600]
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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.
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