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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Polymer Microchips For Capillary Electrophoresis And Electric Field Gradient Focusing Of Biomolecules, Ryan Thomas Kelly Sep 2005

Polymer Microchips For Capillary Electrophoresis And Electric Field Gradient Focusing Of Biomolecules, Ryan Thomas Kelly

Theses and Dissertations

Polymeric materials have seen increasing use as microfluidic device substrates due to their low cost and the simplicity of templated fabrication procedures. I showed that poly(methyl methacrylate) (PMMA) microdevices could be enclosed in a boiling water bath, which allowed the seal to form more quickly than in conventional approaches, and enabled microchannels to remain hydrated throughout the bonding process. Microchip capillary electrophoresis (µ-CE) devices were fabricated using water-based enclosure, and a mixture of fluorescently labeled amino acids was separated in 30 s in these microchips. To create more robust capillary electrophoresis (CE) microdevices with improved separation performance, phase-changing sacrificial ...


Quantitative Nonisotopic Nitrocellulose Filter Binding Assays: Bacterial Manganese Superoxide Dismutase–Dna Interactions, Joshua Czerwinski, Stephanie Hovan, David Mascotti Jan 2005

Quantitative Nonisotopic Nitrocellulose Filter Binding Assays: Bacterial Manganese Superoxide Dismutase–Dna Interactions, Joshua Czerwinski, Stephanie Hovan, David Mascotti

David P. Mascotti

Nitrocellulose filter binding assays (NCFBAs) have been used for many years to qualitatively and quantitatively determine protein–nucleic acid affinities. While this technique can be robust thermodynamically and fairly simple to perform, the requirement of radiolabeled nucleic acids (typically 32P) has several major drawbacks. Some disadvantages are the short half-life of 32P, the inherent safety concerns, and the cost of working with radioisotopes. Another drawback is that over time the beta emissions cause fragmentation of the nucleic acids. We have modified standard NCFBAs by developing a quantitative nonisotopic chemiluminescent method using biotin-labeled DNA and a dual-filter format. The biotin tag ...