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

项目摘要 实现常规DNA测序有可能提高对生物过程的理解， 关系以及在医学，法医学和环境调查方面的援助。然而， 测序的未来好处要求降低成本和速度，并改进仪器 相对于目前商业上可用的技术，在各种下一代DNA中， 随着测序技术的发展，纳米孔链测序已经成为一种非常有前途的方法。 替代.链测序具有作为单分子技术的优点，其能够 长（内切酶）读段，需要最少的样品制备和无样品标记，使用相对 廉价的硬件和生化试剂，对于有效的序列覆盖是高通量的，并且 利用直接读出，具有读取表观遗传修饰和病变以及对其他基因进行测序的潜力。 聚合物因此，为了解决目前需要的改进，以使常规DNA测序通过 使用纳米孔，EBS提出开发方法来实现单核苷酸分辨率，而不使用 通过利用EBS的专有电子器件、αHL孔和新型DNA慢化技术， 机制等这些进步将使得能够开发低复杂性（即不需要PCR）， 高准确度（解析插入缺失和错配）纳米孔链测序方法。

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.