This proposal is to develop a high-throughput, long-read, accurate, fast, and low-cost sequencing method. As the recent next generation sequencing pours out DNA/RNA sequence information at an unprecedented rate, it is becoming apparent that fundamentally these cyclic sequencing methods are unlikely to either reach the long read or produce the accuracy to match the performance of the Sanger method. Meanwhile, the high cost of the current methods, due mainly to large reagent consumptions and the requirement for a high-fold deep coverage, have limited their potential to reach $1,000 genome assembly and to become widely used in traditional sequencing for customized needs for many research and clinical needs. We propose the development of a next generation long-read sequencing technology (NGS, Next Generation Sanger sequencing), which will take advantage of the long time-proven Sanger sequencing and capillary electrophoresis, massively parallel sample preparation, and time-resolved capillary array imaging. On these bases we will establish a new 3D separation system that will provide reliable means to overcome the short-read and stepwise (or cyclic) reaction limitations in all of the present next generation sequencing methods and multiply the throughput of the conventional Sanger sequencing method.
While it is widely understood that decoding of genomic DNA and RNA serves as the foundation for understanding of complex biological systems, it will remain impossible for scientists to unravel the meaning and significance of the myriad of genetic variations and their associations with human health, disease and drug response unless DNA sequencing technology advances to the point that complete and accurate sequence data can be obtained from a much larger population, at a much higher rate and much lower cost than what people have access to today. Pursuant to this goal, this proposed project seeks to overcome the limitations of the traditional and currently commercialized next-generation sequencing technologies. The enabling technology developed herein will likely directly impact medical research in the areas of disease prevention, diagnosis, mechanistic studies and treatment.
| Zhou, Xiaochuan; Zhu, Qi; Eicken, Christoph et al. (2012) MicroRNA profiling using µParaflo microfluidic array technology. Methods Mol Biol 822:153-82 |
