Preparation Of The Polymerase Chain Reaction And Nano-particles On The Interface Of Nanoparticles Aggregates

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. DNA that can easily link functional radical should be attached to nanoparticle surface and amplified largely when PCR proceed. If PCR based on the nanoparticle surface can proceed successfully and be used for the preparation of nanostructure materials, it will be very significant to study in the field of biology,functional material and so on. In this paper, we have systemically studied PCR based on the nanoparticle surface and the preparation of nanostructure aggregates, and obtain several significant results:1. The preparation of nanostructured aggregates using PCR based on nanoparticles was studied. The primers were covalently attached to the nanoparticles surface, and PCR processed using Plasmid pBluescript SK as templates. Atomic Force Microscopy (AFM) and Fluorescence Spectroscopy were used to detect and observe the PCR products. The effects of PCR cycles, size of nanoparticles and length of the amplified DNA fragment were evaluated. The result confirms that nanostructured aggregates can be successfully fabricated when PCR was based on nanoparticles using DNA templates. The nanostructured aggregates formed different shapes and size structures under the control of PCR cycles, size of nanoparticles and length of the amplified DNA fragment, and can be reversed by thermal denaturation Therefore, the aggregates can take on the function of conglomeration and dispersedness with temperature change, when PCR was based on nanoparticles.2. The preparation of dumbbell nanostructured aggregates was studied. First, different size of Au nanoparticles were prepared, which distributing evenly and having uniform particle size. The 5'-SH-(CH2)6- modified primers were covalently attached to the Au nanoparticles (AuNP) surface via Au-S bonds, and PCR was based on AuNP. Nanostructured aggregates were detected by AFM. Optimized conditions of preparing dumbbell nanostructured aggregates were evaluated. The results obtained indicate that PCR can proceed successfully with two kinds of primers respectively bound to the AuNP surface, and dumbbell nanostructured aggregates were prepared. The shapes and size of nanostructured aggregates were effectively controlled through the factors of PCR cycles, size of nanoparticles and length of the amplified DNA fragment.3. PCR based on the Polyamidoamine Dendrimers (PAMAM) surface was studied. The primers were attached to the nanoparticles surface via static electrification, and PCR processed using Plasmid pBluescript SK as templates. Agarose gel electrophoresis and transmission electron microscopes (TEM) were used to detect and observe the PCR products. And the effects of annealing temperature and primers concentration were evaluated, respectively. The results indicate that PCR can proceed successfully under optimized condition with both the two primers respectively bound to the PAMAM surface, and nanostructured aggregates were prepared. In certain temperature scope, PCR has no affections of annealing temperature. The nanostructured aggregates prepared via PCR based on nanoparticles can disperse and congregate reversibly with temperature. Development of PCR procedure based on PAMAM extends its use from liquid homogeneous system to nanoparticle surface. Therefore it not only expands the application range of PCR technology but also offers a novel method to prepare nanostructured material.