Polymerase Chain Reaction Based On The Nanoparticle Surface And Its Application

PCR, a technique for in vitro amplification of a specific nucleic acid fragment, is one of the most important experimental methods in molecular biology. It has been playing an important role in life science, genetics and medicine fields. Usually, PCR is carried out in liquid homogeneous system, whether it could proceed on the surface of nanoparticles or not becomes a challenging problem. If PCR based on the nanoparticle surface can proceed successfully, it will widely expand the applied range of PCR, from liquid homogeneous system to the nanoparticle surface. Therefore it is very significant to study PCR based on the nanoparticle surface. In this paper, we have systemically studied PCR based on the nanoparticle surface and its application, and obtain several significant results:(1) PCR based on the Au nanoparticle (AuNP) surface was studied. The 5'-SH- (CH₂)₆- modified primers were covalently attached to the AuNP surface via Au-S bonds. The linker - (CH₂)₆- between the AuNP surface and primers were used to reduce the steric effect caused by AuNPs. Plasmid pBluescript SK was used as templates. Agarose gel electrophoresis and transmission electron microscope (TEM) were used to detect and observe the PCR products. The effects of the concentration of AuNP-bound primers, annealing temperature and PCR cycle were evaluated, respectively. The results obtained indicate that PCR can proceed successfully with either one kind of primers or two kinds of primers bound to the AuNP surface. More interestingly, AuNPs can be interconnected by dsDNA and nanostructured aggregates are fabricated via PCR with two kinds of primers respectively bound to the AuNP surface.