Browsing by Subject "quantum dots"
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Item An Investigation on Gel Electrophoresis with Quantum Dots End-labeled DNA(2009-05-15) Chen, XiaojiaInvented in the 1950s, gel electrophoresis has now become a routine analytical method to verify the size of nucleic acids and proteins in molecular biology labs. Conventional gel electrophoresis can successfully separate DNA fragments from several base pairs to a few tens of kilo base pairs, beyond which a point is reached that DNA molecules cannot be resolved due to the size independent mobility. In this case, pulsed field gel electrophoresis (PFGE) was introduced to extend the range of DNA fragment sizes that can be effectively separated. But despite the incredible success of PFGE techniques, some important drawbacks remain. First, separation time is extremely long, ranging from several hours to a few days. Second, detection methods still rely on staining the gel after the run. Real time observation and study of band migration behavior is impossible due to the large size of the PFGE device. Finally, many commercial PFGE instruments are relatively expensive, a factor that can limit their accessibility both for routine analytical and preparative use as well as for performing fundamental studies. In this research, a miniaturized PFGE device was constructed with dimension 2cm x 2.6cm, capable of separating DNA fragments ranging from 2.5kb to 32kb within three hours using low voltage. The separation process can be observed in real time under a fluorescence microscope mounted with a cooled CCD camera. Resolution and mobility of the sample were measured to test the efficiency of the device. We also explored manipulating DNA fragments by end labeling DNA molecules with quantum dot nanocrystals. The quantum dot-DNA conjugates can be further modified through binding interactions with biotinylated single-stranded DNA primers. Single molecule visualization was performed during gel electrophoresis and the extension length, entanglement probability and reorientation time of different conjugates were measured to study their effect on DNA migration through the gel. Finally, electrophoresis of DNA conjugates was performed in the miniaturized PFGE device, and shaper bands were observed compared with the non end-labeled sample. Furthermore, by end-labeling DNA with quantum dots, the migration distance of shorter fragments is reduced, providing the possibility of separating a wider range of DNA fragment sizes on the same gel to achieve further device miniaturization.Item Microwave-Assisted Synthesis of II-VI Semiconductor Micro- and Nanoparticles towards Sensor Applications(2013-01-15) Majithia, RavishEngineering particles at the nanoscale demands a high degree of control over process parameters during synthesis. For nanocrystal synthesis, solution-based techniques typically include application of external convective heat. This process often leads to slow heating and allows decomposition of reagents or products over time. Microwave-assisted heating provides faster, localized heating at the molecular level with near instantaneous control over reaction parameters. In this work, microwave-assisted heating has been applied for the synthesis of II-VI semiconductor nanocrystals namely, ZnO nanopods and CdX (X = Se, Te) quantum dots (QDs). Based on factors such as size, surface functionality and charge, optical properties of such nanomaterials can be tuned for application as sensors. ZnO is a direct bandgap semiconductor (3.37 eV) with a large exciton binding energy (60 meV) leading to photoluminescence (PL) at room temperature. A microwave-assisted hydrothermal approach allows the use of sub-5 nm ZnO zero-dimensional nanoparticles as seeds for generation of multi-legged quasi one-dimensional nanopods via heterogeneous nucleation. ZnO nanopods, having individual leg diameters of 13-15 nm and growing along the [0001] direction, can be synthesized in as little as 20 minutes. ZnO nanopods exhibit a broad defect-related PL spanning the visible range with a peak at ~615 nm. Optical sensing based on changes in intensity of the defect PL in response to external environment (e.g., humidity) is demonstrated in this work. Microwave-assisted synthesis was also used for organometallic synthesis of CdX(ZnS) (X = Se, Te) core(shell) QDs. Optical emission of these QDs can be altered ased on their size and can be tailored to specific wavelengths. Further, QDs were incorporated in Enhanced Green-Fluorescent Protein ? Ultrabithorax (EGFP-Ubx) fusion protein for the generation of macroscale composite protein fibers via hierarchal self-assembly. Variations in EGFP- Ubx?QD composite fiber surface morphology and internal QD distribution were studied with respect to (i) time of QD addition (i.e., pre or post protein self-assembly) and (ii) QD surface charge ? negatively charged QDs with dihydrolipoic acid functionalization and positively charged QDs with polyethyleneimine coating. Elucidating design motifs and understanding factors that impact the protein-nanoparticle interaction enables manipulation of the structure and mechanical properties of composite materials.