| Based on the development of current status of Next-Generation Sequencing technologies (NGS), I developed several technologies to improve NGS technologies and exampled to apply the NGS technology to biomedical sciences. In the first chapter, I overviewed most of the NGS technologies, described the characteristics of the technologies, and compared the pits and falls, as well as the applications. Based on the current development status, I developed Dual-Primer emulsion PCR (DPePCR) to simplify the library preparation and elongate the read length, and the details were illustrated in the chapter 2. To minimize the sequencing feature/template and improve sequencing throughput, I developed Rolony sequencing by Synthesis, which was elaborated in chapter 3. To utilize the NGS technologies on biological researches, Haplotype-resolved sequencing was developed to sequence haplotype genome in chapter 4. Haplotype-resolved sequencing technology provided haplotype information, which is very important for fully understanding human genome, completely describing genetic diversities, and identifying the association between genetic variations and diseases. In one word, this dissertation developed several sequencing technologies that improved the next-generation sequencing technologies, and will potentially contribute to biomedical applications. |
