Improved DNA sequencing quality and efficiency using an optimized fast cycle sequencing protocol

摘要

Technological advances in DNA sequencing over the past 20 years have given rise to the science of genomics and have enabled critical advances in other fields, including epidemiology, forensics, evolutionary biology, and medical diagnostics. Further advances in this technology will ensure the proliferation of new applications, such as comparative sequencing, and stimulate a paradigm shift in biology (1). Unfortunately, several factors, primarily the high cost of sequencing reactions, have slowed progress (2). In large genome sequencing centers, reduction of reaction volume along with standardization of template type and quality have contributed to lowered costs (3). Emphasis is now on developing novel sequencing methods to enable genome-wide analysis(4), and these efforts promise to further reduce the cost of large-scale sequencing projects. However, most DNA sequencing projects are of modest scale (5) and still use the dideoxy chain termination method described by Sanger et al. (6). Therefore, improvements to conventional methods that increase efficiency and reduce costs will have immediate benefits to a large number of researchers, especially when implemented in the setting of centralized facilities in which template heterogeneity and variability provide challenges to cost-cutting measures that are effective in genome sequencing centers.