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.

In this paper the manipulation of Ca-alginate microspheres, using a microfluidic chip, for the encapsulation of gold nanoparticles is presented. Our strategy is based on hydrodynamic-focusing on the forming of a series of self-assembling sphere structures, the so-called water-in-oil (w/o) emulsions, in the cross-junction microchannel. These fine emulsions, consisting of aqueous Na-alginates, are then dripped into a solution of 20% calcium salt to accomplish Ca-alginate microspheres in an efficient manner. Experimental data show that microspheres with diameters ranging from 50 µm to 2000 µm with a variation less than 5% were precisely generated. The size and gap of the droplets are tunable by adjusting the relative sheath/sample flow rate ratio. Furthermore, we applied them to encapsulated gold nanoparticles, and this one shot operation performs the ‘Lab on a Chip’.