Achieving Single Nucleotide Resolution to Enable DNA Flossing Through Alpha-Hemolysin

项目总结

Project Summary Enabling routine DNA sequencing has the potential to improve the understanding of biological processes and relationships as well as aid in medicine, forensics, and environmental investigations. However, the potential future benefits of sequencing require a reduction in cost and speed, and an improvement in instrumentation accessibility relative to presently commercially available technology. Among the various next generation DNA sequencing technologies being pursued, nanopore strand sequencing has emerged as a very promising alternative. Strand sequencing has the advantages of being a single-molecule technique that is capable of long (kilobases) reads, requires minimal sample preparation and no sample labeling, uses relatively inexpensive hardware and biochemical reagents, is high-throughput for efficient sequence coverage, and utilizes direct readout with the potential to read epigenetic modifications and lesions as well as sequence other polymers. Thus, to address the current improvements needed to enable routine DNA sequencing through the use of nanopores, EBS proposes to develop methods to achieve single-nucleotide resolution without the use of processive enzymes by taking advantage of EBS’ proprietary electronics, αHL pores and novel DNA slowing mechanisms. These advancements will enable the development of a low complexity (i.e. no PCR needed), high accuracy (indels and mismatches are resolved) nanopore strand sequencing approach.