| Summary: | 碩士 === 國立交通大學 === 應用化學系 === 83 === Separation and detection of DNA restriction fragments are the
essential and major steps in gene analysis or physical mapping
of genes. Slab gel electrophoresis is the trational way to
separate DNA fragments,however, the method is time consuming
,labor intensive, hard to quantitate, and large sample quantity
is required for analysis. To detect DNA fragment in a slab gel
either ethidium bromide or radioisotope is emploted and these
chemicals can pose futher health hazardous problem. Capillary
electrophoresis is an automated separation technique offering
the benefits of rapid and high resolution separation of DNA
molecules. Capillary electrophoresis has been a maturenique for
separating proteins, peptide, organic chemicals, and small
fragments separations, several hurdles imposed by the nature of
large DNA molecules needed to be overcome before the technique
can be realized. In this thesis, I explore several capillary
electrophoresis conditions to separate large DNA restriction
fragments and apply the technique to DNA restriction mapping
which is an essential tool to many molecular biology labs.
Laser induced fluorescence detection and partial digest mapping
method are adopted and modified to map DNA fragments by
capillarytrophoresis. The pros and cons of capillary
electrophoresis in DNA mapping as compared to the tradtional
slab gel electrophoresis are discussed by using pBluescript
SK(+) phagemid DNA as a model. A restriction map of human
interleukine 5 receptor gene is constructed using capillary
electrophoresis and the technique developed in this thesis to
demonstrate and conclude the feasibility of this thesis project.
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