| Summary: | 碩士 === 東海大學 === 化學系 === 101 === The methylation of the promoter region of DNA is an important regulatory mechanism for the downstream gene expression, and the extent of methylation has been linked to cancer formation. In this thesis a simple and cost-effective method was proposed to rapid screening of the DNA methylation by single-strand conformation polymorphism (SSCP) and capillary electrophoresis with laser-induced fluorescence (CE-LIF). In this study, DNA sample was heated at 95 ºC for the denaturation following by cooled at 0 ºC to forms single strand DNA. The different conformation of DNA fragments were then separated by capillary electrophoresis using 1.5% poly(ethylene) oxide (Mave, 8,000,000 Da) in the presence of electroosmotic flow. The electropherograms demonstrated the bisulfite treated, PCR amplified DNA standards (methylated and unmethylated) could be fully separated before 20 minutes at 5 kV. Otherwise, our results also demonstrated that the four hepatocellular carcinoma cells displayed different heterogeneity of DNA methylation. These data also corresponded to the results that obtained from combined bisulfite restriction analysis and DNA sequencing. Therefore, combined SSCP and CE-LIF provided a simple, method for rapid screening of heterogeneity of DNA methylation and may potentially useful for cancer diagnosis or prognosis mornitoring.
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