Impacts of single-walled carbon nanotubes on polymerase chain reaction
Uysal, Ebru (2015) Impacts of single-walled carbon nanotubes on polymerase chain reaction. [Thesis]
Polymerase Chain Reaction (PCR) is an advanced technology used in modern era of molecular biolgy to amplify millions of copies of DNA from a single copy. In order to utilize this technique with its full potential, certain hindrances such as nonspecific by-products, low yield, complexity of GC rich and long genomic DNA amplification need to be eliminated. Among different PCR technologies, Nanomaterial-assisted PCR termed as nanoPCR is a developing technique used to get more improved and the most satisfactory results by using nanomaterials and PCR reaction together. Nanomaterials have been used in PCR due to their unique physical and chemical properties such as high thermal conductivity, stability and high surface to volume ratios that make them significantfor numerous of research areas. The effects of nanomaterials in PCR depend on their size, shape, concentration, heat conductivity, electron transfer properties and surface modifications. Carbon nanotubes have hydrophobic surface area which make them tend to agglomerate into nanoropes in order to minimize surface energy and interaction with the environment. In our study, the effect of Single-Walled Carbon Nanotubes (SWCNTs) was investigated in PCR in order to see their potential as next generation enhancers, where Polyacyrlamide Gel Electrophoresis (PAGE), DynamicLight Scattering (DLS) and Scanning Electron Microscopy (SEM) were mainly performed as core techniques. We demonstrated the impacts of three different SWCNTs in PCR; pristine, amine functionalized and carboxyl functionalized, at their different dispersion states including sonicated, centrifuged and filtered dispersions. The goal was to get rid of agglomerates formed during the reactions. The sonicated single-walled carbon nanotubes are long and have large aggregates as compared to centrifuged and filtered ones which are relatively smaller and short in size. Dynamic Light Scattering (DLS) is used to measure the relative hydrodynamic size of all three; sonicated, centrifuged and filtered single-walled carbon nanotubes to be employed in PCR. Our results showed that intensities of electrophoretic bands of target DNA were affected by introduction of single-walled carbon nanotubes at different dispersion states and concentrations. This work would be useful in the complementary fields like nanobiology, nanomedicine and biosensing.
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