Enhancing the efficiency of a PCR using gold nanoparticles.
We found that the PCR could be dramatically enhanced by Au nanoparticles. With the addition of 0.7 nM of 13 nm Au nanoparticles into the PCR reagent, the PCR efficiency was increased. Especially when maintaining the same or higher amplification yields, the reaction time could be shortened, and the heating/cooling rates could be increased. The excellent heat transfer property of the nanoparticles should be the major factor in improving the PCR efficiency. Different PCR systems, DNA polymerases, DNA sizes and complex samples were compared in this study. Our results demonstrated that Au nanoparticles increase the sensitivity of PCR detection 5- to 10-fold in a slower PCR system (i.e. conventional PCR) and at least 10(4)-fold in a quicker PCR system (i.e. real-time PCR). After the PCR time was shortened by half, the 100 copies/microl DNA were detectable in real-time PCR with gold colloid added, however, at least 10(6) copies/microl of DNA were needed to reach a detectable signal level using the PCR reagent without gold colloid. This innovation could improve the PCR efficiency using non-expensive polymerases, and general PCR reagent. It is a new viewpoint in PCR, that nanoparticles can be used to enhance PCR efficiency and shorten reaction times.
Li
Min
M
Department of Engineering Science, National Cheng Kung University, Tainan, Taiwan, Republic of China.
Lin
Yu-Cheng
YC
Wu
Chao-Chin
CC
Liu
Hsiao-Sheng
HS
D004247:DNA / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D006358:Hot Temperature; D049329:Nanostructures / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
D006046:Gold; D004247:DNA; D004259:DNA-Directed DNA Polymerase