Date of Award

Winter 1996

Project Type

Dissertation

Program or Major

Biochemistry

Degree Name

Doctor of Philosophy

First Advisor

Thomas Laue

Abstract

The analytical electrophoresis apparatus (AEA) is capable of generating and monitoring the electrophoretic migration of macroions. The oligonucleotide pd(A)$\sb{20}\cdot$pd(T)$\sb{20}$ was use as a model to compound to evaluate the range and validity of AEA measurements under a variety of electric fields and solvent conditions. A broad range of electric fields yield consistent, reproducible values. The charge determination from different procedures, steady state electrophoresis (SSE) and electrophoretic mobility, have not been consolidated into a consistent theory but advancements in the scope and understanding of the AEA's potential have been made. The apparent charge from the AEA measured electrophoretic mobility, $\mu$, of Pd(A)$\sb{20}\cdot$pd(T)$\sb{20}$ shows more sensitivity to solution conditions (different chloride salts or ionic strength in a buffer of 20 mM Tris, pH 8.0, at 20$\sp\circ$C) than the SSE determined Q$\sb{\rm app}$. The electrophoretic mobility of pd(A)$\sb{20}\cdot$pd(T)$\sb{20}$ increases with decreasing ionic strength (100 nM KCl to 20 mM KCl salt conditions) and show a direct relationship (possibly coupled flow) to the mobility (and the cation affinity to DNA) of the different chloride salt (Li$\sp+$, Na$\sp+$, K$\sp+$, and (CH$\sb3$)$\sb4$N$\sp{\cdot+}$). The combined SSE data (global nonlinear fit of all fields, one solution condition) suggest a single value for the apparent charge of around 5 e (electron units).

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