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

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 HL 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 Project Narrative Developing a rapid inexpensive and highly accurate means of routine DNA sequencing has the potential to revolutionize numerous fields including medicine forensics and environmental investigations With specific regards to medical applications such advancements could enable more comprehensive and personalized patient screening predictive risk assessment and treatment thus leading to substantial improvements in patient health and survival